Beta-Hydroxy-Gamma-Aminophosphonates and Methods for the Preparation and Use Thereof

ABSTRACT

The present invention provides β-hydroxy-γ-aminophosphonates, β-amino-γ-aminophosphonates, and analogs thereof that inhibit carnitine acyltransferases. The invention also provides compositions comprising these β-hydroxy-γ-aminophosphonates, β-amino-γ-aminophosphonates, and analogs, and methods of the use of such compounds and compositions in the treatment, amelioration or prevention of pathological conditions, diseases or disorders that are linked with fatty acid metabolism, such as non-insulin dependent diabetes or obesity. The invention also provides processes for the preparation of such compounds and compositions.

BACKGROUND OF THE INVENTION

1Field of the Invention

This invention is in the field of medicinal chemistry. In particular,the invention relates to carnitine analogs such asβ-hydroxy-γ-aminophosphonates and β-amino-γ-aminophosphonates thatinhibit carnitine acyltransferases, and intermediates, precursors, andderivatives thereof. In another embodiment, the invention relates to theuse of β-hydroxy-γ-aminophosphonates and β-amino-γ-aminophosphonates,and analogs and derivatives thereof, for the treatment, amelioration orprevention of pathological conditions, diseases or disorders that arelinked with fatty acid metabolism, such as non-insulin dependentdiabetes or obesity. In another embodiment, the invention relates tomethods for the preparation of β-hydroxy-γ-aminophosphonates, andintermediates, precursors, derivatives, and analogs thereof.

2. Related Art

L-carnitine, also known as levocarnitine or vitamin B_(T), is a cofactorthat is present in tissues of animals, including humans, and servesseveral vital physiological roles. In particular, L-carnitine reactswith long chain fatty acids which cannot pass through the mitochondrialmembrane. After such reaction, fatty acids are converted intomembrane-permeable derivatives. In this pathway, L-carnitine plays avital role for the utilization of fatty acids in mitochondria, viaoxidation for the production of energy in eukaryotic organisms. Thiscofactor functions by binding activated fatty acids in the form of acylcarnitine (carnitine shuttle).

The use of L-carnitine in the treatment of hyperlipoproteinemia,hyperlipidemia, and myocardial dysfunction has been the subject ofintense investigation (see, for example, Carazza, U.S. Pat. No.4,255,449; Ramacci, U.S. Pat. No. 4,315,944; Siliprandi, HypolipidemicDrugs, G. Ricci (Ed.), New York; Raven, 1982; Pauly et al., Am. J.Kidney Dis. 41:S35-S43 (2003); Calvani et al., Basic Res. Cardiol.95:75-83 (2000)). L-carnitine has also been reported to be useful as anadjuvant therapy in the management of renal anemia (Ciancuaruso, et al.,Contrib. Nephrol. 137:426-430 (2002)). Certain carnitine analogs orderivatives have also been shown to have potential therapeutic value.For example, propionyl carnitine (the propionic ester of carnitine) hasbeen shown to improve cardiac function (see, for example, Wiseman etal., Drugs Aging 12:243-248 (1998); Ferrari et al., Developments inCardiovascular Medicine 162:323 (1995)). Acetyl carnitine has beenproposed as a possible therapeutic agent for Alzheimer's disease(Pettegrew et al., Expert Review of Neurotherapeutics 2:647-654 (2002)).Bromoacetyl-L-carnitine has been shown in vitro to have a potent effectagainst T. Bruceli, a causative agent of African trypanosomiases(Gilbert et al., Biochem. Pharmacol. 32:3447-3451 (1983)). However, thepotential therapeutic benefit of bromoacetyl-L-carnitine is limitedbecause of toxicity due to metabolic release of bromine and/orbromoacetoacetate.

CPS 124, a carnitine monothiophosphate derivative which is a reversibleand competitive inhibitor of carnitine palmitoyl transferase I, isreportedly undergoing clinical development for the treatment ofnon-insulin dependent diabetes mellitus (NIDDM) (Anderson, Curr. Pharm.Des. 4:1-16 (1998)). Nicotinyl carnitine derivatives have been studiedas anticholesteremics and hypolipemics (Chibata et al., U.S. Pat. No.4,032,641). Acylated aminocarnitines (Griffith, U.S. Pat. No. 4,781,863and Giannessi et al., WO 2008/15081) have been studied asanticholesteremics and hypolipemics.

Carnitine acyltransferases are a group of structurally related enzymesinvolved in lipid catabolism. More specifically, these enzymesparticipate in fatty acid oxidation, catalyzing the exchange of acylgroups between carnitine and Coenzyme A (CoA) (Bieber, Ann. Rev.Biochem. 57:261-283 (1988); Kerner et al., Biochim. Biophys. Acta1486:1-17 (2000); McGarry at al., Eur. J. Biochem. 244:1-14 (1997);Ramsay et al., Biochim. Biophys. Acta 1546:21-43 (2001)). Among thecarnitine acyltransferases are carnitine acetyltransferase (CRAT, alsoknown as CAT), carnitine octanoyltransferase (COT) and carnitinepalmitoyltransferase (CPT), with substrate preferences for short-chain,medium-chain and long-chain fatty acids, respectively. These enzymesgenerally contain approximately 600 amino acid residues and havemolecular weights of about 70 kD. They are the products of a multi-genefamily which may have evolved by duplication of a single ancestral gene(van der Leij et al., Mol. Genet. Metab. 71:139-153 (2000)).

The physiologic relevance of carnitine acyltransferases not only is asource of pathology when these enzymes go awry, but also providesopportunities for treatment of diseases linked with disorders in fattyacid metabolism. The hyperglycemia found in diabetes results fromdecreased glucose disposal concomitant with increased glucoseproduction, which are often associated with increased and uncontrolledfatty acid oxidation (Bebernitz et al., Curr. Pharm. Des. 8:1199-1227(2002)). Hence, inhibition of fatty acid oxidation has emerged as a newstrategy for the treatment of diabetes (Bebernitz et al., Curr. Pharm.Des. 8:1199-1227 (2002); Wagman et al., Curr. Pharm. Des. 7:417-450(2001)), in particular non-insulin dependent diabetes mellitus (“NIDDM”;also known as “mature onset diabetes”).

Thus, there exists a need for carnitine acyltransferase inhibitors forthe treatment of diabetes, obesity, and other diseases that areassociated with disorders in fatty acid metabolism. There also exists aneed for pharmaceutical compositions comprising carnitineacyltransferase inhibitors. There also exists a need for a method ofpreparing carnitine acyltransferase inhibitors.

SUMMARY OF THE INVENTION

The present invention relates to β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates of Formula I and II, or stereoisomers ormixtures of stereoisomers thereof. Compounds of the invention inhibitcarnitine acyltransferases. Carnitine acyltransferase inhibitors haveshown promise in the treatment, amelioration or prevention ofpathological conditions, diseases or disorders that are linked withfatty acid metabolism, including but not limited to, non-insulindependent diabetes mellitus, obesity, hyperlipoproteinemia,hyperlipidemia, cardiac disorders, e.g., myocardial dysfunction, renalanemia, and Alzheimer's disease.

Thus, in one aspect present invention pertains toβ-hydroxy-γ-aminophosphonates and β-amino-γ-aminophosphonates of FormulaI:

particularly wherein:

-   R is selected from the group consisting of hydrogen and lower alkyl;-   R¹ is selected from the group consisting of hydrogen, optionally    substituted alkyl, aralkyl, optionally substituted cycloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted aryl, optionally substituted heteroaryl, and    optionally substituted heterocyclo;-   R² and R³ are independently selected from the group consisting of    hydrogen, optionally substituted alkyl, aralkyl, optionally    substituted aryl, and monovalent pharmaceutically acceptable cation,    or taken together R² and R³ represent a divalent pharmaceutically    acceptable cation;-   R⁴ is selected from the group consisting of hydrogen, optionally    substituted alkyl, optionally substituted cycloalkyl, optionally    substituted alkenyl, optionally substituted alkynyl, and COR⁵;-   R⁵ is selected from the group consisting of optionally substituted    alkyl, aralkyl, optionally substituted cycloalkyl, optionally    substituted alkenyl, optionally substituted alkynyl, optionally    substituted aryl, optionally substituted heteroaryl, and optionally    substituted heterocyclo;-   X⁻ is a pharmaceutically acceptable anion, or X⁻ and R² are absent    and the compound of Formula I is a zwitterion;-   Z is selected from the group consisting of O and NR¹⁰; and-   R¹⁰ is selected from the group consisting of hydrogen, optionally    substituted alkyl, aralkyl, optionally substituted cycloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted aryl, optionally substituted heteroaryl, and    optionally substituted heterocyclo;-   or pharmaceutically acceptable hydrates, crystalline forms or    amorphous forms thereof, or a stereoisomer or mixture of    stereoisomers thereof.

In additional embodiments, the β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates of the present invention are compoundsselected from the group consisting of Formulae 3S-I; 3R-I; 2R,3S-I;2S,3S-I; 2R,3R-I; and 2S,3R-I:

or pharmaceutically acceptable hydrates, crystalline forms or amorphousforms of anyone thereof, wherein R, R¹, R², R³, R⁴, X⁻ and Z have themeanings as described above for Formula I.

In additional embodiments, the β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates of the present invention are zwitterioniccompounds of Formula II:

or pharmaceutically acceptable hydrates, crystalline forms or amorphousforms thereof, wherein R , R¹, R³, R⁴ and Z have the meanings asdescribed above for Formula I.

In additional embodiments, the β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates of the present invention are zwitterioniccompounds selected from the group consisting of Formulae 3S-II; 3R-II;2R,3S-II; 2S,3S-II; 3R,3R-II; and 2S,3R-II:

or pharmaceutically acceptable hydrates, crystalline forms or amorphousforms of anyone thereof, wherein R, R¹, R³, R⁴, and Z have the meaningsas described above for Formula I.

The present invention also provides methods for the preparation of aβ-hydroxy-γ-aminophosphonate of Formula III:

particularly wherein:

-   R is selected from the group consisting of hydrogen and lower alkyl;-   R¹ is selected from the group consisting of hydrogen, optionally    substituted alkyl, aralkyl, optionally substituted cycloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted aryl, optionally substituted heteroaryl, and    optionally substituted heterocyclo;-   R^(2a) and R^(3a) are independently selected from the group    consisting of optionally substituted alkyl, aralkyl, and optionally    substituted aryl; and-   X⁻ is a pharmaceutically acceptable anion;-   the methods comprising:-   (a) reacting a compound of Formula IV

with RX, to give a compound of Formula III, or a stereoisomer or mixtureof stereoisomers thereof; and

-   (b) isolating said compound of Formula III.

In one such embodiment, a compound of Formula III is a diastereomericmixture having S-stereochemistry at the 3-position, i.e., a compound ofFormula 3S-III:

In another such embodiment, a compound of Formula III is adiastereomeric mixture having R-stereochemistry at the 3-position, i.e.,a compound of Formula 3R-III:

In still another such embodiment, a compound of Formula III is in the2R,3S-isomeric form, i.e., a compound of Formula 2R,3S-III:

In still another such embodiment, a compound of Formula III is in the2S, 3S-isomeric form, i.e., a compound of Formula 2S,3S-III:

In still another such embodiment, a compound of Formula III is in the2R,3R-isomeric form, i.e., a compound of Formula 2R,3R-III:

In still another such embodiment, a compound of Formula III is the2S,3R-isomer, i.e., a compound of Formula 2R,3R-III:

The present invention also provides methods for separating thestereoisomers, e.g., diastereomers, of a compound of Formula III. Thus,in one embodiment, the present invention provides methods for thepreparation of a compound of Formula 2S,3S-III from a compound ofFormula 3S-III, comprising isolating a compound of Formula 2S,3S-III,i.e., the 2S,3S-isomer, substantially free from a compound of Formula2R,3S-III, i.e., the 2R,3S-isomer. In another embodiment, the inventionprovides methods for the preparation of a compound of Formula 2R,3S-IIIfrom a compound of Formula 3S-III, comprising isolating a compound ofFormula 2R,3S-III substantially free from a compound of Formula2S,3S-III. In another embodiment, the invention provides methods for thepreparation of a compound of Formula 2R,3R-III from a compound ofFormula 3R-III, comprising isolating a compound of Formula 2R,3R-III,i.e., the 2R,3R-isomer, substantially free from a compound of Formula2S,3R-III, i.e., the 2S,3R-isomer. In another embodiment, the inventionprovides methods for the preparation of a compound of Formula 2S,3R-IIIfrom a compound of Formula 3R-III, comprising isolating a compound ofFormula 2S,3R-III substantially free from a compound of Formula2R,3R-III.

The present invention also provides methods for the preparation of acompound of

Formula IV, the methods comprising:

-   (a) removing R⁶, or removing R⁶ and R⁷, from a compound of Formula    V:

particularly wherein:

-   R⁶ is an amine protecting group; and-   R⁷ is selected from the group consisting of hydrogen and an amine    protecting group, or R⁶ and R⁷ taken together represent an amine    protecting group; and R¹, R^(2a) and R^(3a) have the meanings as    described above for Formula III, to give a compound of Formula IV,    or a stereoisomer or mixture of stereoisomers thereof; and-   (b) isolating said compound of Formula IV; or-   (c) using said compound of Formula IV in the next reaction without    isolation.

In one such embodiment, a compound of Formula V is a diastereomericmixture having S-stereochemistry at the 3-position, i.e. a compound ofFormula 3S-V:

In another such embodiment, a compound of Formula V is a diastereomericmixture having R-stereochemistry at the 3-position, i.e. a compound ofFormula 3R-V:

The present invention also provides methods for the preparation of aβ-hydroxy-γ-aminophosphonate of Formula V; the methods comprising:

-   (a) reducing a β-keto-γ-aminophosphonate of Formula VI:

particularly wherein R¹, R^(2a) and R^(3a) have the meanings asdescribed above for Formula III, and R⁶ and R⁷ have the meanings asdescribed above for Formula V, to give a compound of Formula VI, or astereoisomer or mixture of stereoisomers thereof; and

-   (b) isolating said compound of Formula V; or-   (c) using said compound of Formula V in the next reaction without    isolation.

In one such embodiment, a compound of Formula VI has S-stereochemistryat the 3-position, i.e. a compound of Formula 3S-VI:

In another such embodiment, a compound of Formula VI hasR-stereochemistry at the 3-position, i.e. a compound of Formula 3R-VI:

The present invention also provides methods for the preparation of acompound of Formula VI, the methods comprising:

-   (a) condensing a compound of Formula VII:

with a compound of Formula VIII:

particularly wherein R¹, R^(2a) and R^(3a) have the meanings asdescribed above for Formula III, R⁶ and R⁷ have the meanings asdescribed above for Formula V, and R⁸ is selected from the groupconsisting of optionally substituted alkyl, aralkyl, and optionallysubstituted aryl, to give a compound of Formula VI, or a stereoisomer ormixture of stereoisomers thereof; and

-   (b) isolating said compound of Formula VI; or-   (c) using said compound of Formula VI in the next reaction without    isolation.

In one such embodiment, a compound of Formula VII is the S-isomer, i.e.a compound of Formula S-VII:

In another such embodiment, a compound of Formula VII is the R-isomer,i.e. a compound of Formula R-VII:

The present invention also provides methods for the preparation of acompound of Formula VIII, the methods comprising:

-   (a) condensing a compound of Formula IX:

with LiR⁹, particularly wherein R^(2a) and R^(3a) have the meanings asdescribed above for Formula III, and R⁹ is selected from the groupconsisting of lower alkyl, and aryl, to give a compound of Formula VIII;and

-   (b) isolating said compound of Formula VIII; or-   (c) using said compound of Formula VIII in the next reaction without    isolation.

The present invention also provides methods for the preparation of acompound of Formula X:

particularly wherein R and R¹ have the meanings as described above forFormula III, R^(2b) and R^(3b) are selected from the group consisting ofhydrogen and monovalent pharmaceutically acceptable cation, or takentogether R^(2b) and R^(3b) represent a divalent pharmaceuticallyacceptable cation, or X⁻ and R^(2b) are absent (i.e., a compound ofFormula X is a zwitterion), the methods comprising:

-   (a) removing R^(2a) and R^(3a) from a compound of Formula III, to    give a compound of Formula X, or a stereoisomer or mixture of    stereoisomers thereof; and-   (b) isolating said compound of Formula X.

In one such embodiment, a compound of Formula X is a diastereomericmixture having S-stereochemistry at the 3-position, i.e., a compound ofFormula 3S-X:

In another such embodiment, a compound of Formula X is a diastereomericmixture having R-stereochemistry at the 3-position, i.e., a compound ofFormula 3R-X.

In still another such embodiment, a compound of Formula X is in the2R,3S-isomeric form, i.e., a compound of Formula 2R,3S-X:

In still another such embodiment, a compound of Formula X is in the2S,3S-isomeric form, i.e., a compound of Formula 2S,3S-X:

In still another such embodiment, a compound of Formula X is in the2R,3R-isomeric form, i.e., a compound of Formula 2R,3R-X:

In still another such embodiment, a compound of Formula X is in the2S,3R-isomeric form, i.e., a compound of Formula 2S,3R-X:

The invention also provides methods for the preparation of a compound ofFormula VII, the method comprising:

-   (a) protecting the amine of an amino acid of Formula XI;-   (b) esterifying the carboxylic acid of an amino acid of Formula XI:

particularly wherein R¹ has the meaning as described above for FormulaIII, to give a compound of Formula VII; and

-   (c) isolating said compound of Formula VII; or-   (d) using said compound of Formula VII in the next reaction without    isolation.

In one such embodiment, the compound of Formula XI is the S-isomerhaving Formula S-XI:

In another such embodiment, the compound of Formula XI is the R-isomerhaving Formula R-XI:

The invention also provides compounds prepared in accordance with themethods of the invention.

The invention also provides pharmaceutical compositions comprising acompound of the invention and a pharmaceutically acceptable carrier.

The invention also provides methods for inhibiting carnitineacyltransferase in a cell comprising contacting said cell with acompound of the invention.

The invention also provides a method of treating, ameliorating, orpreventing a disorder or condition responsive to the inhibition ofcarnitine acyltransferase in a patient in need thereof, comprisingadministering to the patient a therapeutically effective amount of acompound of the invention.

The invention also provides a method of treating, ameliorating, orpreventing a disorder or condition responsive to the inhibition ofcarnitine acyltransferase in a patient in need thereof, comprisingadministering to the patient a therapeutically effective amount of apharmaceutical composition comprising a compound of the invention andone or more additional therapeutic agents.

The invention also provides a method of treating, ameliorating, orpreventing a disorder or condition responsive to the inhibition ofcarnitine acyltransferase in a patient in need thereof, comprisingadministering to the patient a therapeutically effective amount of acompound of the invention in combination with one or more additionaltherapeutic agents.

Other features and advantages of the invention will be apparent from thefollowing detailed description, and from the claims. The disclosedmaterials, methods, and examples are for illustrative purposes only andare not intended to be limiting. Skilled artisans will appreciate thatmethods and materials similar or equivalent to those described hereincan be used to practice the invention.

Unless otherwise defined, all technical and scientific terms used hereinhave the meaning commonly understood by one skilled in the art to whichthis invention belongs. All publications, patent applications, patents,and other references mentioned herein are incorporated by reference intheir entirety. In case of conflict, the present specification,including definitions, will control.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a bar graph showing extracellular glucose levels measured inconditioned media of hepatic cells treated with different concentrationsof the diastereomeric mixture of compound 7.

FIG. 2 is a bar graph showing intracellular glucose levels in hepaticcells treated with different concentrations of the diastereomericmixture of compound 7.

FIG. 3 is a bar graph showing intracellular glycogen levels in hepaticcells treated with different concentrations of the diastereomericmixture of compound 7.

FIG. 4 is a bar graph showing the effect of the diastereomeric mixtureof compound 7 on cell viability of renal and hepatic cells.

FIG. 5 is a line graph showing the effect of the diastereomeric mixtureof compound 7 on cell proliferation of renal cells.

FIG. 6 is a line graph showing the effect of the diastereomeric mixtureof compound 7 on cell proliferation of hepatic cells.

FIG. 7 is a series of five illustrations that show the morphologicalappearance of chick embryos treated with different concentrations of thediastereomeric mixture of compound 7.

DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS

The term “alkyl” as used herein by itself or part of another grouprefers to a straight-chain or branched saturated aliphatic hydrocarbonhaving from one to eighteen carbons or the number of carbons designated,e.g., C₁-C₁₈ means from 1 to 18 carbons, inclusive. In one suchembodiment, the alkyl is a C₁-C₁₀ alkyl. In another such embodiment, thealkyl is a C₁-C₈ alkyl. In certain such embodiments, the alkyl is alower alkyl. Non-limiting exemplary alkyl groups according to certainaspects of the invention include methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl,isohexyl, n-heptyl, 4,4-dimethylpentyl, n-octyl, 2,2,4-trimethylpentyl,nonyl, decyl, and the like. Other suitable alkyl groups will be familiarto those of ordinary skill in the relevant arts.

The term “lower alkyl” as used herein by itself or part of another groupmeans the alkyl as defined above has 1 to 6 carbons, i.e., aC₁-C₆-alkyl. Non-limiting exemplary lower alkyl groups include methyl,ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl,n-pentyl, isopentyl, n-hexyl, and the like. Other suitable lower alkylgroups will be familiar to those of ordinary skill in the relevant arts.

The term “optionally substituted alkyl” as used herein by itself or partof another group means that the alkyl as defined above is eitherunsubstituted or substituted with one, two or three substituentsindependently selected from hydroxy, i.e., —OH, nitro, i.e., —NO₂,cyano, i.e., —CN, halo, amino, optionally substituted cycloalkyl,optionally substituted heteroaryl, optionally substituted heterocyclo,alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. Inone such embodiment, the optionally substituted alkyl is unsubstituted.In another such embodiment, the optionally substituted alkyl issubstituted with one substituent. In another such embodiment, theoptionally substituted alkyl is substituted with two substituents. Incertain such embodiments, the substituents are selected from hydroxy,i.e., a hydroxyalkyl, halo, i.e., a haloalkyl, or amino, i.e., anaminoalkyl. In certain such embodiments, the optionally substitutedalkyl is an optionally substituted C₁-C₆-alkyl, i.e., an optionallysubstituted lower alkyl. Exemplary optionally substituted alkyl groupsinclude, but are not limited to, —CH₂OCH₃, —CH₂CH₂NH₂, —CH₂CH₂CN,—CH₂CONH₂, hydroxymethyl, hydroxyethyl, hydroxypropyl, trifluoromethyl,and the like. Other suitable optionally substituted alkyl groups will befamiliar to those of ordinary skill in the relevant arts.

The term “aralkyl” as used herein by itself or part of another grouprefers to an optionally substituted alkyl as defined above having one,two or three optionally substituted aryl substituents. In one suchembodiment, the optionally substituted alkyl is unsubstituted. Inanother such embodiment, the optionally substituted aryl isunsubstituted. In certain such embodiments, the optionally substitutedaryl is phenyl (abbreviated as “Ph”). In another such embodiment, thearalkyl has one optionally substituted aryl substituent. In another suchembodiment, the aralkyl has two optionally substituted arylsubstituents. In a particular embodiment, the aralkyl is an aryl(C₁-C₄alkyl). In certain such embodiments, the aryl(C₁-C₄ alkyl) has oneoptionally substituted aryl substituent. Non-limiting exemplary aralkylgroups include, for example, benzyl, phenylethyl, (4-fluorophenyl)ethyl,phenylpropyl, diphenylmethyl (i.e., Ph₂CH—), diphenylethyl (i.e.,Ph₂CHCH₂—), and the like. Other suitable aralkyl groups will be familiarto those of ordinary skill in the relevant arts.

The term “cycloalkyl” as used herein by itself or part of another grouprefers to saturated and partially unsaturated (containing one or twodouble bonds) cyclic hydrocarbon groups containing one to three ringshaving from three to twelve carbon atoms (i.e., C₃-C₁₂ cycloalkyl) orthe number of carbons designated. In one such embodiment, the cycloalkylhas one ring. In another such embodiment, the cycloalkyl is a C₃-C₇cycloalkyl. Non-limiting exemplary cycloalkyl groups includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, norbornyl, decalin, adamantyl, and the like. Other suitablecycloalkyl groups will be familiar to those of ordinary skill in therelevant arts.

The term “optionally substituted cycloalkyl” as used herein by itself orpart of another group means the cycloalkyl as defined above is eitherunsubstituted or substituted with one, two or three substituentsindependently selected from halo, nitro, cyano, hydroxy, amino,optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl,optionally substituted cycloalkyl, optionally substituted alkenyl,optionally substituted alkynyl, optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted heterocyclo, alkoxy,aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. The term“optionally substituted cycloalkyl” also means the cycloalkyl as definedabove may be fused to an optionally substituted aryl. Non-limitingexemplary optionally substituted cycloalkyl groups include:

and the like. Other suitable optionally substituted cycloalkyl groupssuitable for use in accordance with this aspect of the invention will befamiliar to those of ordinary skill in the relevant arts.

The term “alkenyl” as used herein by itself or part of another grouprefers to an alkyl group as defined above containing one, two or threecarbon-to-carbon double bonds. In one embodiment, the alkenyl has onecarbon-to-carbon double bond. Non-limiting exemplary alkenyl groupsinclude —CH═CH₂, —CH₂CH═CH₂, —CH₂CH₂CH═CH₂, —CH₂CH₂CH═CHCH₃ and thelike. Other suitable alkenyl groups will be familiar to those ofordinary skill in the relevant arts.

The term “optionally substituted alkenyl” as used herein by itself orpart of another group means the alkenyl as defined above is eitherunsubstituted or substituted with one, two or three substituentsindependently selected from halo, nitro, cyano, hydroxy, amino,optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl,optionally substituted cycloalkyl, optionally substituted alkenyl,optionally substituted alkynyl, optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted heterocyclo, alkoxy,aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. Non-limitingexemplary optionally substituted alkenyl groups include —CH═CH—,—CH═CHPh, —CH₂CH═CHPh, and the like. Other suitable optionally alkenylgroups will be familiar to those of ordinary skill in the relevant arts.

The term “alkynyl” as used herein by itself or part of another grouprefers to an alkyl group as defined above containing one to threecarbon-to-carbon triple bonds. In one embodiment, the alkynyl has onecarbon-to-carbon triple bond. Non-limiting exemplary alkynyl groupsinclude —C≡CH, —C≡CCH₃, —CH₂C≡CH, —CH₂CH₂C≡CH and —CH₂CH₂CCCH₃. Othersuitable alkynyl groups will be familiar to those of ordinary skill inthe relevant arts.

The term “optionally substituted alkynyl” as used herein by itself orpart of another group means the alkynyl as defined above is eitherunsubstituted or substituted with one, two or three substituentsindependently selected from halo, nitro, cyano, hydroxy, amino,optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl,optionally substituted cycloalkyl, optionally substituted alkenyl,optionally substituted alkynyl, optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted heterocyclo, alkoxy,aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. Non-limitingexemplary optionally substituted alkenyl groups include —C≡CPh,—CH₂C≡CPh and the like. Other suitable optionally substituted alkynylgroups will be familiar to those of ordinary skill in the relevant arts.

The term “aryl” as used herein by itself or part of another group refersto monocyclic and bicyclic aromatic ring systems having from six tofourteen carbon atoms (i.e., C₆-C₁₄ aryl) such as phenyl (abbreviated asPh), 1-naphthyl and 2-naphthyl, and the like. Other aryl groups suitablefor use in accordance with this aspect of the invention will be familiarto those of ordinary skill in the relevant arts.

The term “optionally substituted aryl” as used herein by itself or partof another group means the aryl as defined above is either unsubstitutedor substituted with one to five substituents independently selected fromhalo, nitro, cyano, hydroxy, amino, optionally substituted alkyl,haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, optionally substituted heteroaryl,optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy,alkylthio, carboxamido or sulfonamido. In one such embodiment, theoptionally substituted aryl is an optionally substituted phenyl, whichin certain embodiments has four substituents, three substituents, twosubstituents or one substituent. Non-limiting exemplary substituted arylgroups include 2-methylphenyl, 2-methoxyphenyl, 2-fluorophenyl,2-chlorophenyl, 2-bromophenyl, 3-methylphenyl, 3-methoxyphenyl,3-fluorophenyl, 3-chlorophenyl, 4-methylphenyl, 4-ethylphenyl,4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 2,6-di-fluorophenyl,2,6-di-chlorophenyl, 2-methyl, 3-methoxyphenyl, 2-ethyl,3-methoxyphenyl, 3,4-di-methoxyphenyl, 3,5-di-fluorophenyl3,5-di-methylphenyl and 3,5-dimethoxy, 4-methylphenyl and the like. Asused herein, the term “optionally substituted aryl” is also meant toinclude groups having fused optionally substituted cycloalkyl and fusedoptionally substituted heterocyclo rings. Non-limiting exemplaryexamples include:

and the like. Additional suitable optionally substituted aryl groups foruse in accordance with this aspect of the invention will be familiar tothose of ordinary skill in the relevant arts.

The term “heteroaryl” as used herein by itself or part of another grouprefers to monocyclic and bicyclic aromatic ring systems typically havingfrom five to fourteen carbon atoms (i.e., C₅-C₁₄ heteroaryl) and one,two, three or four heteroatoms independently selected from the groupconsisting of oxygen, nitrogen and sulfur. In one such embodiment, theheteroaryl has four heteroatoms. In another such embodiment, theheteroaryl has three heteroatoms. In another such embodiment, theheteroaryl has two heteroatoms. In another such embodiment, theheteroaryl has one heteroatom. Non-limiting exemplary heteroaryl groupsinclude 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-imidazolyl, 4-imidazolyl,pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridylpyrimidyl, 4-pyrimidyl, purinyl, 2-benzimidazolyl, 4-benzimidazolyl,5-benzimidazolyl, 2-benzthiazolyl, 4-benzthiazolyl, 5-benzthiazolyl,5-indolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 1-isoquinolyl,5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 2-quinolyl, 3-quinolyl,6-quinolyl and the like. As used herein, the term “heteroaryl” is alsomeant to include possible N-oxides. Non-limiting exemplary N-oxidesinclude pyridyl N-oxide and the like. Additional suitable heteroarylgroups for use in accordance with this aspect of the invention will befamiliar to those of ordinary skill in the relevant arts.

The term “optionally substituted heteroaryl” as used herein by itself orpart of another group means the heteroaryl as defined above is eitherunsubstituted or substituted with one to four substituents, typicallyone or two substituents, which are typically independently selected fromhalo, nitro, cyano, hydroxy, amino, optionally substituted alkyl,haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, optionally substituted heteroaryl,optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy,alkylthio, carboxamido or sulfonamido. In one such embodiment, theoptionally substituted heteroaryl has one substituent. According to thisaspect of the invention, any available carbon or nitrogen atom may besubstituted. Non-limiting exemplary optionally substituted heteroarylgroups include:

and the like. Additional suitable optionally substituted heteroarylgroups for use in accordance with this aspect of the invention will befamiliar to those of ordinary skill in the relevant arts.

The term “heterocyclo” as used herein by itself or part of another grouprefers to saturated and partially unsaturated (containing one or twodouble bonds) cyclic groups containing one to three rings having fromtwo to twelve carbon atoms (i.e., C₂-C₁₂ heterocyclo) and one or twooxygen, sulfur or nitrogen atoms. According to this aspect of theinvention, the heterocyclo can be optionally linked to the rest of themolecule through a carbon or nitrogen atom. Non-limiting exemplaryheterocyclo groups include:

and the like. Additional suitable heterocyclo groups for use inaccordance with this aspect of the invention will be familiar to thoseof ordinary skill in the relevant arts.

The term “optionally substituted heterocyclo” as used herein by itselfor part of another group means the heterocyclo as defined above iseither unsubstituted or substituted with one to four substituents whichare typically independently selected from halo, nitro, cyano, hydroxy,amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl,optionally substituted cycloalkyl, optionally substituted alkenyl,optionally substituted alkynyl, optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted heterocyclo, alkoxy,aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. Substitutionmay occur on any available carbon or nitrogen atom. Non-limitingexemplary substituted heterocyclo groups include:

and the like. In certain embodiments of the invention, an optionallysubstituted heterocyclo may be fused to an aryl group to provide anoptionally substituted aryl as described above.

The term “alkoxy” as used herein by itself or part of another grouprefers to a haloalkyl, optionally substituted alkyl, optionallysubstituted cycloalkyl, optionally substituted alkenyl or optionallysubstituted alkynyl attached to a terminal oxygen atom. Non-limitingexemplary alkoxy groups include methoxy, tert-butoxy, —OCH₂CH═CH₂ andthe like.

The term “aryloxy” as used herein by itself or part of another grouprefers to an optionally substituted aryl attached to a terminal oxygenatom. Non-limiting exemplary aryloxy groups include phenoxy and thelike.

The term “aralkyloxy” as used herein by itself or part of another grouprefers to an aralkyl attached to a terminal oxygen atom. Non-limitingexemplary aralkyloxy groups include benzyloxy and the like.

The term “alkylthio” as used herein by itself or part of another grouprefers to a haloalkyl, aralkyl, optionally substituted alkyl, optionallysubstituted cycloalkyl, optionally substituted alkenyl or optionallysubstituted alkynyl attached to a terminal sulfur atom. Non-limitingexemplary alkyl groups include —SCH₃ and the like.

The term “halo” or “halogen” as used herein by itself or part of anothergroup refers to fluoro, chloro, bromo or iodo. In certain embodiments ofthe present invention, the halo is fluoro or chloro.

The term “amino” as used herein by itself or part of another grouprefers to a radical of formula —NR^(a)R^(b) wherein R^(a) and R^(b) areindependently hydrogen, haloalkyl, aralkyl, optionally substitutedalkyl, optionally substituted cycloalkyl, optionally substitutedheterocyclo, optionally substituted aryl or optionally substitutedheteroaryl; or R^(a) and R^(b) taken together with the nitrogen atom towhich they are attached form a four to seven membered optionallysubstituted heterocyclo. Non-limiting exemplary amino groups include—NH₂, —N(H)CH₃, —N(CH₃)₂, N(H)CH₂CH₃, N(CH₂CH₃), —N(H)CH₂Ph and thelike.

The term “carboxamido” as used herein by itself or part of another grouprefers to a radical of formula —CO-amino. Non-limiting exemplarycarboxamido groups include —CONH₂, —CON(H)CH₃, —CON(H)Ph,—CON(H)CH₂CH₂Ph, —CON(CH₃)₂, CON(H)CHPh₂ and the like.

The term “sulfonamido” as used herein by itself or part of another grouprefers to a radical of formula —SO₂-amino. Non-limiting exemplarysulfonamido groups include —SO₂NH₂, —SO₂N(H)CH₃, —SO₂N(H)Ph and thelike.

The term “about,” as used herein, includes the recited number ±10%.Thus, “about 10” means 9 to 11, inclusive.

The term “leaving group” as used herein refers to an atom or group thatbecomes detached from an atom or group in what is considered to be theresidual or main part of the substrate in a specified reaction. In amidecoupling reactions, exemplary leaving groups (i.e., leaving groupsdesignated L¹) include —F, —Cl, —Br, —OH, —OC₆F₅, —O(CO)alkyl and thelike. In one embodiment, the leaving group, L¹, is —Cl. In anotherembodiment, the leaving group, L¹, is an activated form of —OH (e.g.,OBt, O-acylisourea). In certain such embodiments of the invention, anactivating agent (e.g., dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC),benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate(PyBop)) may be employed to active a carboxylic acid (i.e., the leavinggroup is —OH) toward amide formation. Such activating agents are wellknown to those of skill in the art of organic synthesis. Otheradditives, such as N-hydroxybenzotriazole (HOBt) or N-hydroxysuccinimide(HOSu), may also be added to optimize reaction parameters (e.g., rate,yield, purity, racemization). In nucleophilic displacement reactions(e.g., S_(N)2 reactions), exemplary leaving groups (i.e., leaving groupsdesignated L²) include Cl, —Br, —I, —OSO₂Me (mesylate), —OSO₂CF₃(triflate), OSO₂C₆H₅ (besylate), —OSO₂CH₃C₆H₄ (tosylate) and the like.In one embodiment, the leaving group, L², is —OSO₂CF₃ or —I In anotherembodiment, the leaving group, L², is —OSO₂CF₃. In another embodiment,the leaving group L² is —I.

The term “amine protecting group” as used herein refers to group thatblocks (i.e., protects) the amine functionality while reactions arecarried out on other functional groups or parts of the molecule. Thoseskilled in the art will be familiar with the selection, attachment, andcleavage of amine protecting groups and will appreciate that manydifferent protective groups are known in the art, the suitability of oneprotective group or another being dependent on the particular thesynthetic scheme planned. Treatises on the subject are available forconsultation, such as Greene and Wuts, “Protective Groups in OrganicSynthesis,” 3rd Ed., pp. 17-245 (J. Wiley & Sons, 1999), the disclosureof which is incorporated herein by reference. Suitable amine protectinggroups include the carbobenzyloxy (Cbz), tert-butyloxycarbonyl (BOC),9-fluorenylmethyloxycarbonyl (FMOC), phthalimide and benzyl (Bn) groups.With regard to a compound of Formula VII, in one such embodiment, theamine protecting group, R⁶, is the carbobenzyloxy ortert-butyloxycarbonyl, and R⁷ is hydrogen. In another such embodiment,the amine protecting groups, R⁶ and R⁷, are benzyl. In another suchembodiment, R⁶ and R⁷ taken together form an amine protecting group,such as a phthalimide group.

The term “C₁-C₄ alcohol” as used herein refers to an alcohol having 1 to4 carbons such as methanol, ethanol, propanol, isopropanol, butanol,isobutanol, sec-butanol and tert-butanol. In one embodiment, the C₁-C₄alcohol is methanol.

The term “monovalent pharmaceutically acceptable cation” as used hereinrefers to inorganic cations such as, but not limited to, alkaline metalions, e.g., Na⁺ and K⁻, as well as organic cations such as, but notlimited to, ammonium and substituted ammonium ions, e.g., NH₄ ⁺, NHMe₃⁺, NH₂Me₂ ⁺, NHMe₃ ⁺ and NMe₄ ⁺.

The term “divalent pharmaceutically acceptable cation” as used hereinrefers to inorganic cations such as, but not limited to, alkaline earthmetal cations, e.g., Ca²⁺ and Mg²⁺.

Examples of monvalent and divalent pharmaceutically acceptable cationsare discussed in Berge et al. J. Pharm. Sci., 66, 1997, 1-19, thedisclosure of which is incorporated herein by reference.

The term “pharmaceutically acceptable anion” as used herein refers to ananion associated with a quaternary ammonium compound of the presentinvention that is acceptable for administration to a patient, e.g., amammal, e.g., a human. In one embodiment, the pharmaceuticallyacceptable anion is the anion of a pharmaceutically acceptable inorganicacid, e.g., hydrochloric, perchloric, sulfuric, phosphoric, hydrobromic,hydroiodic or nitric acid and the like. In one embodiment, thepharmaceutically acceptable anion is the anion of a pharmaceuticallyacceptable organic acid, e.g., a mono or polyvalent organic acid, e.g.,citric, fumaric, maleic, malic, ascorbic, succinic, tartaric, benzoic,acetic, phenylacetic, methanesulfonic, ethansulfonic, benzenesulfonic orp-toluenesulfonic acid and the like.

The term “pharmaceutically acceptable salt,” as used herein, refers toany salt (e.g., obtained by reaction with an acid or a base) of acompound of the present invention that is physiologically tolerated inthe target animal (e.g., a mammal, such as a human). Salts of thecompounds of the present invention may be derived from inorganic ororganic acids and bases. Examples of suitable acids include, but are notlimited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric,fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic,toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic,ethanesulfonic, formic, benzoic, boronic, malonic, sulfonic, picolinic,naphthalene-2-sulfonic, benzenesulfonic acid, and the like. Other acids,such as oxalic, while not in themselves pharmaceutically acceptable, maybe employed in the preparation of salts useful as intermediates inobtaining the compounds of the invention and their pharmaceuticallyacceptable acid addition salts.

Examples of suitable bases include, but are not limited to, alkali metal(e.g., sodium) hydroxides, alkaline earth metal (e.g., magnesium)hydroxides, ammonia, and compounds of formula NW₄ ⁺, wherein W is C₁₋₄alkyl, and the like.

Examples of suitable such salts include, but are not limited to:acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,bisulfate, borate, boronate, butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate,hemisulfate, heptanoate, hexanoate, chloride, bromide, iodide,2-hydroxyethanesulfonate, lactate, maleate, mesylate, methanesulfonate,2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate,persulfate, phenylpropionate, picrate, pivalate, propionate, succinate,tartrate, thiocyanate, tosylate, undecanoate, nitrate, sulfate,picolinate, besylate, perchloriate, salicylate, phosphate, and the like.Other examples of suitable salts according to the invention includeanions of the compounds of the present invention compounded with asuitable cation such as Na⁺, K₊, Ca²⁺, Mg²⁺, Mn²⁺, NH₄ ⁺, and NW₄ ⁺(wherein W is a C₁₋₄ alkyl group), and the like, including additionalpharmaceutically acceptable salts that are well known in the art (see,e.g., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pa., 19th ed. 1995) and others that are known to those of ordinary skillin the relevant arts. For therapeutic use, salts of the compounds of thepresent invention are contemplated as being pharmaceutically acceptable.However, salts of acids and bases that are non-pharmaceuticallyacceptable may also find use, for example, in the preparation orpurification of a pharmaceutically acceptable compound.

The term “one pot process” as used herein refers to a strategy toimprove the efficiency of a compound synthesis whereby a reactant issubjected to successive chemical reactions in one reactor or reactionvessel. Use of this process potentially avoids the need for lengthyseparation processes and purification of the synthetic intermediates. Inaddition, a one pot process may increase chemical yield. The chemicalconversion of leucine to(3-trimethylamonium-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethylester; iodide without isloation and/or purification of any syntheticintermediates is an example of a one pot process.

The term “pharmaceutical composition” as used herein refers to acomposition comprising one or more active pharmaceutical ingredientsincluding, but not limited to, one or more compounds of the inventionwhich can be used to treat, prevent or reduce the severity of a disease,disorder or condition in a subject, e.g., a mammal such as a human, thatis suffering from, that is predisposed to, or that has been exposed tothe disease, disorder or condition. A pharmaceutical compositiongenerally comprises an effective amount of one or more active agents,e.g., a compound of the present invention such as a compound of FormulaI or II, or a stereoisomer or mixture of stereoisomers thereof, and apharmaceutically acceptable carrier. The pharmaceutical composition canalso a comprise a compound of the invention and one or more additionalingredients, including but not limited to one or more therapeutic agentssuch as one or more anticholesterolemics, one or more statins, e.g.,atorvastatin, lovastatin, pravastatin, rosuvastatin, fluvastatin orsimvastatin, one or more anticoagulants, e.g., warfarin, one or moreanti-obesity drugs, e.g., orlistat or sibutramine, or one or moreanti-diabetic drugs, such as one or more sulfonylureas, e.g.,glimepiride, glibenclamide, one or more biguanides, e.g., metformin, orone or more glitazones, e.g., pioglitazone or rosiglitazone.

The term “pharmaceutically acceptable carrier” encompasses any of thestandard pharmaceutical carriers, buffers and excipients, includingphosphate-buffered saline solution, water, and emulsions (such as anoil/water or water/oil emulsion), and various types of wetting agentsand/or adjuvants. Suitable pharmaceutical carriers and theirformulations are described in Remington's Pharmaceutical Sciences, MackPublishing Co., Easton, Pa., 19th ed. 1995. Preferred pharmaceuticalcarriers depend upon the intended mode of administration of the activeagent. Typical modes of administration are described below.

The term “therapeutically effective amount,” as used herein, refers tothat amount of a given therapeutic agent sufficient to result inamelioration of one or more symptoms of a disorder or condition, orprevent appearance or advancement of a disorder or condition, or causeregression of or cure from the disorder or condition.

The term “therapeutic agent,” as used herein refers to any chemicalsubstance that can be used in the treatment, management, prevention oramelioration of a disease, condition or disorder or one or more symptomsthereof. Suitable therapeutic agents include, but are not limited to,small molecules, synthetic drugs, peptides, polypeptides, proteins,nucleic acids (e.g., DNA and RNA polynucleotides including, but notlimited to, antisense nucleotide sequences, triple helices, andnucleotide sequences encoding biologically active proteins,polypeptides, or peptides), antibodies, synthetic or natural inorganicmolecules, mimetic agents, and synthetic or natural organic molecules.In some embodiments, the therapeutic agent is one which is known to beuseful for, or has been or is currently being used for, the treatment,management, prevention or amelioration of a condition or disorder or oneor more symptoms thereof.

The term “borohydride reducing agent” as used herein refers to aborohydride-based reducing agent capable of reducing a ketone to asecondary alcohol, e.g., sodium borohydride, lithium borohydride,borane, etc. In one embodiment, the borohydride reducing agent is sodiumborohydride.

The stereochemical terms and conventions used in the specification areconsistent with those described in Pure & Appl. Chem 68:2193 (1996),unless otherwise indicated.

The term “purity,” as used herein, refers to chemical and/orstereoisomeric (i.e., diastereomeric or enantiomeric) purity, unlessotherwise indicated.

The term “enantiomeric excess” or “ee” refers to a measure for how muchof one enantiomer is present compared to the other. For a mixture of Rand S enantiomers, the percent enantiomeric excess is defined as|R−S|*100, where R and S are the respective mole or weight fractions ofenantiomers in a mixture such that R+S=1. With knowledge of the opticalrotation of a chiral substance, the percent enantiomeric excess isdefined as ([α]_(obs)/[α]_(max))*100, where [α]_(obs) is the opticalrotation of the mixture of enantiomers and [α]_(max) is the opticalrotation of the pure enantiomer.

The term “diastereomeric excess” or “de” refers to a measure for howmuch of one diastereomer is present compared to the other and is definedby analogy to enantiomeric excess. Thus, for a mixture of diastereomers,D1 and D2, the percent diastereomeric excess is defined as |D1−D2|*100,where D1 and D2 are the respective mole or weight fractions ofdiastereomers in a mixture such that D1+D2=1.

The determination of diastereomeric and/or enantiomeric excess ispossible using a variety of analytical techniques, including NMRspectroscopy, chiral column chromatography and/or optical polarimetryaccording to routine protocols will be familiar to those of ordinaryskill in the art.

The term “substantially free,” as used herein, refers to a compositioncomprising at least about 90% by weight of one stereoisomer, i.e.,enantiomer or diastereomer, over the corresponding stereoisomer. Inanother embodiment, the composition comprises at least about 91%, about92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%,about 99% or about 99.5% by weight of the desired stereoisomer. Thus,the term “a compound of Formula 2S,3S-III substantially free from acompound of Formula 2R,3S-III,” as used herein, refers to a compositioncomprising at least about 90% of a compound of Formula 2S,3S-III and atmost about 10% of a compound of Formula 2R,3S-III. Similarly, the term“a compound of Formula 2R,3S-III substantially free from a compound ofFormula 2S,3S-III,” as used herein, refers to a composition comprisingat least about 90% of a compound of Formula 2R,3S-III and at most about10% of a compound of Formula 2S,3S-III.

The term “anti-solvent” as used herein refers to a solvent that reducesthe solubility of a solute, e.g., a compound of Formula III, in asolution thereby facilitating precipitation, i.e., crystal growth. Inone embodiment, the anti-solvent is selected from the group consistingof hexane, ethyl acetate, acetone, methyl ethyl ketone and methylt-butyl ether.

Throughout the specification, groups and optional substituents thereofare chosen to provide stable moieties and compounds.

Compounds of the present invention exist as stereoisomers includingoptical isomers. The invention includes all stereoisomers, as pureindividual stereoisomer preparations and as enriched preparations ofeach, and as the racemic mixtures of such stereoisomers as well as theindividual enantiomers and diastereomers that may be separated accordingto methods that are well-known to those of skill in the art.

Overview

The present invention provides β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates, or stereoisomers or mixtures ofstereoisomers thereof, that inhibit carnitine acyltransferases. Thus,compounds of the invention are useful for the treatment, amelioration orprevention of pathological conditions, diseases or disorders that arelinked with fatty acid metabolism, including but not limited to,non-insulin dependent diabetes mellitus, obesity, hyperlipoproteinemia,hyperlipidemia, cardiac disorders, e.g., myocardial dysfunction, renalanemia, and Alzheimer's disease. The present invention also providespharmaceutical compositions comprising one or more compounds of theinvention, and optionally, one or more additional therapeutic agents.The present invention also provides methods of treating, ameliorating,or preventing a disorder or condition responsive to the inhibition ofcarnitine acyltransferase in a patient, comprising administering to thepatient a therapeutically effective amount of a compound of theinvention, and optionally one or more additional therapeutic agents. Thepresent invention also provides methods for the preparation ofβ-hydroxy-γ-aminophosphonates and β-amino-γ-aminophosphonates, orstereoisomers or mixtures of stereoisomers thereof.

Thus, in one embodiment, the present invention providesβ-hydroxy-γ-aminophosphonates and β-amino-γ-aminophosphonates of FormulaI:

particularly wherein:

-   R is selected from the group consisting of hydrogen and lower alkyl;-   R¹ is selected from the group consisting of hydrogen, optionally    substituted alkyl, aralkyl, optionally substituted cycloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted aryl, optionally substituted heteroaryl, and    optionally substituted heterocyclo;-   R² and R³ are independently selected from the group consisting of    hydrogen, optionally substituted alkyl, aralkyl, optionally    substituted aryl, and monovalent pharmaceutically acceptable cation,    or taken together R² and R³ represent a divalent pharmaceutically    acceptable cation;-   R⁴ is selected from the group consisting of hydrogen, optionally    substituted alkyl, optionally substituted cycloalkyl, optionally    substituted alkenyl, optionally substituted alkynyl, and COR⁵;-   R⁵ is selected from the group consisting of optionally substituted    alkyl, aralkyl, optionally substituted cycloalkyl, optionally    substituted alkenyl, optionally substituted alkynyl, optionally    substituted aryl, optionally substituted heteroaryl, and optionally    substituted heterocyclo;-   X⁻ is a pharmaceutically acceptable anion, or X⁻ and R² are absent    and the compound of Formula I is a zwitterion,-   Z is selected from the group consisting of O and NR¹⁰; and-   R¹⁰ is selected from the group consisting of hydrogen, optionally    substituted alkyl, aralkyl, optionally substituted cycloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted aryl, optionally substituted heteroaryl, and    optionally substituted heterocyclo;-   or pharmaceutically acceptable hydrates, crystalline forms or    amorphous forms thereof, or a stereoisomer or mixture of    stereoisomers thereof.

In one embodiment, a compound of Formula I is a mixture ofstereoisomers, e.g., a mixture of diastereomers and/or enantiomers. Inanother embodiment, a compound of Formula I is a mixture ofdiastereomers. In another embodiment, a compound of Formula I is amixture of enantiomers. In another embodiment, a compound of Formula Iis a single enantiomer.

In another particular embodiment, β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonate analogs of the present invention arecompounds selected from the group consisting of Formula 3S-I; 3R-I;2R,3S-I;, 2S,3S-I; 2R,3R-I; and 2S,3R-I:

or pharmaceutically acceptable hydrates, crystalline forms or amorphousforms of any one thereof, particularly wherein R, R¹, R², R³, R⁴, X⁻ andZ have the meanings as described above for Formula I.

In one embodiment, β-hydroxy-γ-aminophosphonate orβ-amino-γ-aminophosphonate of the present invention is a compound ofFormula 3S-I, i.e., a diastereomeric mixture having S-stereochemistry atthe 3-position, i.e., the γ-position. In another embodiment, aβ-hydroxy-γ-aminophosphonate or β-amino-γ-aminophosphonate analog of thepresent invention is a compound of Formula 3R-I, i.e., a diastereomericmixture having R-stereochemistry at the 3-position. In anotherembodiment, a β-hydroxy-γ-aminophosphonate or β-amino-γ-aminophosphonateanalog of the present invention is a compound of Formula 2R,3S-I, i.e.,the 2R,3S-isomer. In another embodiment, a β-hydroxy-γ-aminophosphonateor β-amino-γ-aminophosphonate analog of the present invention is acompound of Formula 2S,3S-I, i.e., the 2S,3S-isomer. In anotherembodiment, a β-hydroxy-γ-aminophosphonate or β-amino-γ-aminophosphonateanalog of the present invention is a compound of Formula 2R,3R-I, i.e.,the 2R,3R-isomer. In another embodiment, a β-hydroxy-γ-aminophosphonateor β-amino-γ-aminophosphonate analog of the present invention is acompound of Formula 2S,3R-I, i.e., the 2S,3R-isomer.

In certain such embodiments, a compound of Formula I, or a stereoisomeror mixture of stereoisomers thereof, has purity of about 90% or more,e.g., about 91%, about 92%, about 93%, about 94%, about 95%, about 96%,about 97%, about 98%, about 99%, about 99.5% or more.

As it relates to a compound of Formula I, or a stereoisomer or a mixtureof stereoisomers thereof, e.g. , a compound of Formulae 3S -I; 3R-I;2R,3S -I; 2S ,3S -I; 2R,3R-I; or 2S,3R-I; in one embodiment, R is loweralkyl. In another such embodiment, R is methyl.

In additional embodiments, R¹ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl, and optionallysubstituted aryl. In additional embodiments, R¹ is selected from thegroup consisting of lower alkyl, aralkyl, and optionally substitutedaryl. In certain such embodiments, R¹ is selected from the groupconsisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, benzyl, and phenyl. In another embodiment, R¹ isisobutyl.

In additional embodiments, R² and R³ are independently selected from thegroup consisting of hydrogen, optionally substituted lower alkyl,aralkyl and optionally substituted aryl. In certain such embodiments, R²and R³ are independently selected from the group consisting of hydrogen,lower alkyl, aralkyl, and aryl. In certain such embodiments, R² and R³are independently selected from the group consisting of hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, isopenyl, benzyl, and phenyl. In additional embodiments, R²and R³ are independently selected from the group consisting of hydrogenand methyl. In another embodiment, R² and R³ are hydrogen.

In another embodiment, R² is a monovalent pharmaceutically acceptablecation and R³ is selected from the group consisting of hydrogen,optionally substituted alkyl, aralkyl, and optionally substituted aryl.In one such embodiment, the monovalent pharmaceutically acceptablecation is selected from the group consisting of Na⁺ and K⁺.

In additional embodiments, R² and R³ are each a monovalentpharmaceutically acceptable cation. In one such embodiment, themonovalent pharmaceutically acceptable cation is selected from the groupconsisting of Na⁺ and K⁺.

In additional embodiments, R² and R³ taken together represent a divalentpharmaceutically acceptable cation. In one such embodiment, the divalentpharmaceutically acceptable cation is selected from the group consistingof Mg²⁺ and Ca²⁺.

In additional embodiments, R⁴ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl, and COR⁵. In one suchembodiment, R⁴ is hydrogen. In another such embodiment, R⁴ is optionallysubstituted lower alkyl. In still another such embodiment, R⁴ is loweralkyl. In additional such embodiments, R⁴ is COR⁵. In one suchembodiment, R⁵ is optionally substituted lower alkyl. In another suchembodiment, R⁵ is lower alkyl, such as methyl, ethyl, propyl or butyl.

In additional embodiments, X⁻ is a pharmaceutically acceptable anionwhich in certain embodiments is selected from the group consisting ofhydroxide, chloride, bromide, iodide, sulphate, nitrate, phosphate,formate, acetate, maleate, fumarate, citrate, oxalate, succinate,tartrate, malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate. In one suchembodiment, X⁻ is a pharmaceutically acceptable anion selected from thegroup consisting of hydroxide, chloride, bromide and iodide.

In additional embodiments, X⁻ and R² are absent and the compound ofFormula I is a zwitterion.

In additional embodiments, Z is NR¹⁰. In one such embodiment, R¹⁰ isselected from the group consisting of hydrogen and optionallysubstituted lower alkyl, particularly hydrogen.

In additional embodiments, Z is O.

In additional embodiments, R is lower alkyl and R¹ is selected from thegroup consisting of optionally substituted lower alkyl, aralkyl, andoptionally substituted aryl. In one such embodiment, R is methyl and R¹is selected from the group consisting of lower alkyl, aralkyl, andoptionally substituted aryl. In another such embodiment, R is methyl andR¹ is selected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl.In another such embodiment, R is methyl and R¹ is isobutyl.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of optionally substituted lower alkyl, aralkyl, andoptionally substituted aryl, and R² and R³ are independently selectedfrom the group consisting of hydrogen, optionally substituted loweralkyl, aralkyl, and optionally substituted aryl. In one such embodiment,R is methyl, R¹ is selected from the group consisting of optionallysubstituted lower alkyl, aralkyl, and optionally substituted aryl, andR² and R³ are independently selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl, and optionallysubstituted aryl. In another such embodiment, R is methyl, R¹ isselected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,and R² and R³ are independently selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl and phenyl. In another suchembodiment, R is methyl, R¹ is isobutyl and R² and R³ are selected fromthe group consisting of hydrogen or methyl. In another such embodiment,R is methyl, R¹ is isobutyl and R² and R³ are hydrogen.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of lower alkyl, aralkyl, and optionally substitutedaryl, R² and R³ are independently selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl, and optionallysubstituted aryl, and R⁴ is selected from the group consisting ofhydrogen and COR⁵. In one such embodiment, R is lower alkyl, R¹ isselected from the group consisting of lower alkyl, aralkyl, andoptionally substituted aryl, R² and R³ are independently selected fromthe group consisting of hydrogen, optionally substituted lower alkyl,aralkyl, and optionally substituted aryl, and R⁴ is hydrogen. In anothersuch embodiment, R is methyl, R¹ is selected from the group consistingof lower alkyl, aralkyl, and optionally substituted aryl, R² and R³ areindependently selected from the group consisting of hydrogen, optionallysubstituted lower alkyl, aralkyl, and optionally substituted aryl, andR⁴ is selected from the group consisting of hydrogen and COR⁵. Inanother such embodiment, R is methyl, R¹ is selected from the groupconsisting of lower alkyl, aralkyl, and optionally substituted aryl, R²and R³ are independently selected from the group consisting of hydrogen,optionally substituted lower alkyl, aralkyl and optionally substitutedaryl, and R⁴ is hydrogen. In another such embodiment, R is methyl, R¹ isselected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,R² and R³ are independently selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl, and R⁴ is selectedfrom the group consisting of hydrogen and COR⁵. In still another suchembodiment, R is methyl, R¹ is selected from the group consisting ofmethyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, benzyl and phenyl, R² and R³ are independently selected fromthe group consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl,and R⁴ is hydrogen. In another such embodiment, R is methyl, R¹ isisobutyl, R² and R³ are selected from the group consisting of hydrogenor methyl, and R⁴ is selected from the group consisting of hydrogen andCOR⁵. In another such embodiment, R is methyl, R¹ is isobutyl, R² and R³are hydrogen, and R⁴ is selected from the group consisting of hydrogenand COR⁵. In another such embodiment, R is methyl, R¹ is isobutyl, R²and R³ are hydrogen, and R⁴ is hydrogen.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of lower alkyl, aralkyl and optionally substitutedaryl, R² and R³ are independently selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl and optionallysubstituted aryl, R⁴ is selected from the group consisting of hydrogenand COR⁵, Z is NR¹⁰, R¹⁰ is selected from the group consisting ofhydrogen and optionally substituted lower alkyl, and X⁻ is apharmaceutically acceptable cation, which in certain embodiments isselected from the group consisting of hydroxide, chloride, bromide,iodide, sulphate, nitrate, phosphate, formate, acetate, maleate,fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate,lactate, gluconate, trifluoroacetate, methanesulphonate, besylate andp-toluenesulphonate. In one such embodiment, R is lower alkyl, R¹ isselected from the group consisting of lower alkyl, aralkyl andoptionally substituted aryl, R² and R³ are independently selected fromthe group consisting of hydrogen, optionally substituted lower alkyl,aralkyl and optionally substituted aryl, R⁴ and R¹⁰ are hydrogen, and X⁻is selected from the group consisting of hydroxide, chloride, bromide,iodide, sulphate, nitrate, phosphate, formate, acetate, maleate,fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate,lactate, gluconate, trifluoroacetate, methanesulphonate, besylate, andp-toluenesulphonate. In another such embodiment, R is methyl, R¹ isselected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,R² and R³ are independently selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl, R⁴ is selected fromthe group consisting of hydrogen and COR⁵, R¹⁰ is hydrogen, and X⁻ isselected from the group consisting of hydroxide, chloride, bromide,iodide, sulphate, nitrate, phosphate, formate, acetate, maleate,fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate,lactate, gluconate, trifluoroacetate, methanesulphonate, besylate andp-toluenesulphonate. In still another such embodiment, R is methyl, R¹is selected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,R² and R³ are independently selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl, R⁴ and R¹⁰ arehydrogen, and X⁻ is selected from the group consisting of hydroxide,chloride, bromide, iodide, sulphate, nitrate, phosphate, formate,acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate,malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate. In another suchembodiment, R is methyl, R¹ isobutyl, R² and R³ are independentlyselected from the group consisting of hydrogen and methyl, R⁴ isselected from the group consisting of hydrogen and COR⁵, R¹⁰ ishydrogen, and X⁻ is selected from the group consisting of hydroxide,chloride, bromide, iodide, sulphate, nitrate, phosphate, formate,acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate,malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate. In still anothersuch embodiment, R is methyl, R¹ isobutyl, R² and R³ are independentlyselected from the group consisting of hydrogen and methyl, R⁴ and R¹⁰are hydrogen, and X⁻ is selected from the group consisting of hydroxide,chloride, bromide, iodide, sulphate, nitrate, phosphate, formate,acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate,malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate. In another suchembodiment, R is methyl, R¹ isobutyl, R² and R³ are hydrogen, R⁴ and R¹⁰are hydrogen, and X⁻ is selected from the group consisting of hydroxide,chloride, bromide, iodide, sulphate, nitrate, phosphate, formate,acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate,malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of lower alkyl, aralkyl and optionally substitutedaryl, R² and R³ are independently selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl and optionallysubstituted aryl, R⁴ is selected from the group consisting of hydrogenand COR⁵, Z is O, and X⁻ is a pharmaceutically acceptable cation, whichin certain embodiments is selected from the group consisting ofhydroxide, chloride, bromide, iodide, sulphate, nitrate, phosphate,formate, acetate, maleate, fumarate, citrate, oxalate, succinate,tartrate, malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate. In one suchembodiment, R is lower alkyl, R¹ is selected from the group consistingof lower alkyl, aralkyl and optionally substituted aryl, R² and R³ areindependently selected from the group consisting of hydrogen, optionallysubstituted lower alkyl, aralkyl and optionally substituted aryl, R⁴ ishydrogen, Z is O, and X⁻ is selected from the group consisting ofhydroxide, chloride, bromide, iodide, sulphate, nitrate, phosphate,formate, acetate, maleate, fumarate, citrate, oxalate, succinate,tartrate, malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate, and p-toluenesulphonate. In another suchembodiment, R is methyl, R¹ is selected from the group consisting ofmethyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, benzyl, and phenyl, R² and R³ are independently selectedfrom the group consisting of hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl, benzyl,and phenyl, R⁴ is selected from the group consisting of hydrogen andCOR⁵, Z is O, and X⁻ is selected from the group consisting of hydroxide,chloride, bromide, iodide, sulphate, nitrate, phosphate, formate,acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate,malate, mandelate, lactate, gluconate, trifluoroacetate,methanesulphonate, besylate and p-toluenesulphonate. In still anothersuch embodiment, R is methyl, R¹ is selected from the group consistingof methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, benzyl, and phenyl, R² and R³ are independently selectedfrom the group consisting of hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl, benzyl,and phenyl, R⁴ is hydrogen, Z is O, and X⁻ is selected from the groupconsisting of hydroxide, chloride, bromide, iodide, sulphate, nitrate,phosphate, formate, acetate, maleate, fumarate, citrate, oxalate,succinate, tartrate, malate, mandelate, lactate, gluconate,trifluoroacetate, methanesulphonate, besylate and p-toluenesulphonate.In another such embodiment, R is methyl, R¹ isobutyl, R² and R³ areindependently selected from the group consisting of hydrogen and methyl,R⁴ is selected from the group consisting of hydrogen and COR⁵, Z is O,and X⁻ is selected from the group consisting of hydroxide, chloride,bromide, iodide, sulphate, nitrate, phosphate, formate, acetate,maleate, fumarate, citrate, oxalate, succinate, tartrate, malate,mandelate, lactate, gluconate, trifluoroacetate, methanesulphonate,besylate and p-toluenesulphonate. In still another such embodiment, R ismethyl, R¹ isobutyl, R² and R³ are independently selected from the groupconsisting of hydrogen and methyl, R⁴ is hydrogen, Z is O, and X⁻ isselected from the group consisting of hydroxide, chloride, bromide,iodide, sulphate, nitrate, phosphate, formate, acetate, maleate,fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate,lactate, gluconate, trifluoroacetate, methanesulphonate, besylate andp-toluenesulphonate. In another such embodiment, R is methyl, R¹isobutyl, R² and R³ are hydrogen, R⁴ is hydrogen, Z is O, and X⁻ isselected from the group consisting of hydroxide, chloride, bromide,iodide, sulphate, nitrate, phosphate, formate, acetate, maleate,fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate,lactate, gluconate, trifluoroacetate, methanesulphonate, besylate andp-toluenesulphonate.

In additional embodiments, β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates of the present invention are zwitterioniccompounds of Formula II:

or pharmaceutically acceptable hydrates, crystalline forms or amorphousforms thereof, particularly wherein R, R¹, R³, R⁴ and Z have themeanings as described above for Formula I.

In one such embodiment, a compound of Formula II is a mixture ofstereoisomers, e.g., a mixture of diastereomers and/or enantiomers.Thus, in one particular such embodiment, a compound of Formula II is amixture of diastereomers. In another particular such embodiment, acompound of Formula I is a mixture of enantiomers. In yet anotherembodiment, a compound of Formula II is a single enantiomer.

In additional embodiments, the β-hydroxy-γ-aminophosphonates andβ-amino-γ-aminophosphonates of the present invention are zwitterioniccompounds of Formulae 3S-II; 3R-II; 2R,3S-II; 2S,3S-II; 3R,3R-II; or2S,3R-II:

or pharmaceutically acceptable hydrates, crystalline forms or amorphousforms of any one thereof, particularly wherein R, R¹, R³, R⁴ and Z havethe meanings as described above for Formula I.

In one such embodiment, a β-hydroxy-γ-aminophosphonate orβ-amino-γ-aminophosphonate analog is a zwitterionic compound of Formula3S-II, i.e., a diastereomeric mixture having S-stereochemistry at the3-position. In another such embodiment, a β-hydroxy-γ-aminophosphonateor β-amino-γ-aminophosphonate analog is a zwitterionic compound ofFormula 3R-II, i.e., a diastereomeric mixture having R-stereochemistryat the 3-position. In another such embodiment, aβ-hydroxy-γ-aminophosphonate or β-amino-γ-aminophosphonate analog is azwitterionic compound of Formula 2R,3S-II, i.e., the 2R, 3S-isomer. Inanother such embodiment, a β-hydroxy-γ-aminophosphonate orβ-hydroxy-γ-aminophosphonate analog is a zwitterionic compound ofFormula 2S,3S-II, i.e., the 2S,3S-isomer. In another embodiment, aβ-hydroxy-γ-aminophosphonate or β-amino-γ-aminophosphonate analog is azwitterionic compound of Formula 2R,3R-II, i.e., the 2R,3R-isomer. Inanother embodiment, the β-hydroxy-γ-aminophosphonate orβ-amino-γ-aminophosphonate analog is a zwitterionic compound of Formula2S,3R-II, i.e., the 2S,3R-isomer.

In certain such embodiments, a compound of Formula II, or a stereoisomeror mixture of stereoisomers thereof, has a purity of about 90% or more,e.g., about 90%, about 91%, about 92%, about 93%, about 94%, about 95%,about 96%, about 97%, about 98%, about 99%, about 99.5% or more.

As it relates to a compound of Formula II, or a stereoisomer or mixtureof stereoisomers thereof, e.g., a compound of Formulae 3S-II; 3R-II;2R,3S-II; 2S,3S-II; 3R,3R-II or 2S,3R-II, in one embodiment, R is loweralkyl. In another such embodiment, R is methyl.

In additional embodiments, R¹ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl and optionallysubstituted aryl.

In additional embodiments, R¹ is selected from the group consisting oflower alkyl, aralkyl, and optionally substituted aryl. In one suchembodiment, R¹ is selected from the group consisting of methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl,and phenyl, and particularly isobutyl.

In additional embodiments, R³ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl, and optionallysubstituted aryl.

In additional embodiments, R³ is selected from the group consisting ofhydrogen, lower alkyl, aralkyl, and aryl. In one such embodiment, R³ isselected from the group consisting of hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl, benzyl,and phenyl, particularly hydrogen or methyl, and more particularlyhydrogen.

In additional embodiments, R⁴ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl and COR⁵. In one suchembodiment, R⁴ is hydrogen. In another such embodiment, R⁴ is optionallysubstituted lower alkyl. In one embodiment, R⁴ is lower alkyl. In stillanother such embodiment, R⁴ is COR⁵. In one such embodiment, R⁵ isoptionally substituted lower alkyl. In another such embodiment, R⁵ islower alkyl, such as methyl.

In additional embodiments, Z is NR¹⁰. In one such embodiment, R¹⁰ isselected from the group consisting of hydrogen and optionallysubstituted alkyl. In another such embodiment, R¹⁰ is hydrogen.

In additional embodiments, Z is O.

In additional embodiments, R is lower alkyl and R¹ is selected from thegroup consisting of optionally substituted lower alkyl, aralkyl, andoptionally substituted aryl. In one such embodiment, R is methyl and R¹is selected from the group consisting of lower alkyl, aralkyl, andoptionally substituted aryl. In another such embodiment, R is methyl andR¹ is selected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl.In another such embodiment, R is methyl and R¹ is isobutyl.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of lower alkyl, aralkyl and optionally substitutedaryl, and R³ is selected from the group consisting of hydrogen,optionally substituted lower alkyl, aralkyl and optionally substitutedaryl. In one such embodiment, R is methyl, R¹ is selected from the groupconsisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, benzyl, and phenyl, and R³ is selected from thegroup consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl.In another such embodiment, R is methyl, R¹ is isobutyl, and R³ isselected from the group consisting of hydrogen or methyl, particularlyhydrogen.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of lower alkyl, aralkyl, and optionally substitutedaryl, R³ is selected from the group consisting of hydrogen, optionallysubstituted lower alkyl, aralkyl, and optionally substituted aryl, R⁴ isselected from the group consisting of hydrogen and COR⁵, Z is NR¹⁰, andR¹⁰ is selected from the group consisting of hydrogen and optionallysubstituted lower alkyl. In one such embodiment, R is lower alkyl, R¹ isselected from the group consisting of lower alkyl, aralkyl, andoptionally substituted aryl, R³ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl, and optionallysubstituted aryl, Z in NR¹⁰, and R⁴ and R¹⁰ are hydrogen. In anothersuch embodiment, R is methyl, R¹ is selected from the group consistingof methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, benzyl, and phenyl, R³ is selected from the group consistingof hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl, Z is NR¹⁰, R¹⁰ ishydrogen, and R⁴ is selected from the group consisting of hydrogen andCOR⁵. In such another embodiment, R is methyl, R¹ is selected from thegroup consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl, R³ is selected fromthe group consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl,and R⁴ is hydrogen. In another such embodiment, R is methyl, R¹ isisobutyl, R³ is selected from the group consisting of hydrogen ormethyl, Z is NR¹⁰, R¹⁰ is hydrogen, and R⁴ is selected from the groupconsisting of hydrogen and COR⁵. In another such embodiment, R ismethyl, R¹ is isobutyl, R³ is hydrogen, Z is NR¹⁰, R¹⁰ is hydrogen, andR⁴ is selected from the group consisting of hydrogen and COR⁵. In oneparticular embodiment, R is methyl, R¹ is isobutyl, Z is NR¹⁰, and R³,R⁴, and R¹⁰ are hydrogen.

In additional embodiments, R is lower alkyl, R¹ is selected from thegroup consisting of lower alkyl, aralkyl, and optionally substitutedaryl, R³ is selected from the group consisting of hydrogen, optionallysubstituted lower alkyl, aralkyl, and optionally substituted aryl, R⁴ isselected from the group consisting of hydrogen and COR⁵, and Z is O. Inone such embodiment, R is lower alkyl, R¹ is selected from the groupconsisting of lower alkyl, aralkyl, and optionally substituted aryl, R³is selected from the group consisting of hydrogen, optionallysubstituted lower alkyl, aralkyl, and optionally substituted aryl, Z isO, and R⁴ is hydrogen. In another such embodiment, R is methyl, R¹ isselected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,R³ is selected from the group consisting of hydrogen, methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl,benzyl, and phenyl, Z is O, and R⁴ is selected from the group consistingof hydrogen and COR⁵. In such another embodiment, R is methyl, R¹ isselected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,R³ is selected from the group consisting of hydrogen, methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl,benzyl, and phenyl, Z is O, and R⁴ is hydrogen. In another suchembodiment, R is methyl, R¹ is isobutyl, R³ is selected from the groupconsisting of hydrogen or methyl, Z is O, and R⁴ is selected from thegroup consisting of hydrogen and COR⁵. In another such embodiment, R ismethyl, R¹ is isobutyl, R³ is hydrogen, Z is O, and R⁴ is selected fromthe group consisting of hydrogen and COR⁵. In one particular embodiment,R is methyl, R¹ is isobutyl, Z is O, and R³ and R⁴ are hydrogen.

In certain embodiments of the invention the compound is selected fromthe group consisting of:

or pharmaceutically acceptable salts, esters, amides, zwitterions orother pharmaceutically acceptable derivatives, variants or formsthereof.

In another such embodiment, the compound is selected from the groupconsisting of:

or pharmaceutically acceptable salts, esters, zwitterions or otherpharmaceutically acceptable derivatives, variants or forms thereof.

In another such embodiment, the compound is:

or a pharmaceutically acceptable salt, ester, zwitterion or otherpharmaceutically acceptable derivative, variant or form thereof.

Methods of Synthesis

In additional embodiments, the present invention also provides methodsfor the preparation of a β-hydroxy-γ-aminophosphonate of Formula III:

particularly wherein:

-   R is selected from the group consisting of hydrogen and lower alkyl;-   R¹ is selected from the group consisting of hydrogen, optionally    substituted alkyl, aralkyl, optionally substituted cycloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted aryl, optionally substituted heteroaryl, and    optionally substituted heterocyclo;-   R^(2a) and R^(3a) are independently selected from the group    consisting of optionally substituted alkyl, aralkyl, and optionally    substituted aryl; and-   X⁻ is a pharmaceutically acceptable anion, the methods comprising:-   (a) reacting a compound of Formula IV

with RX, to give a compound of Formula III, or a stereoisomer or mixtureof stereoisomers thereof; and

-   (b) isolating said compound of Formula III.

In one such embodiment, a compound of Formula III is a diastereomericmixture having S-stereochemistry at the 3-position, referred to as the“3S-isomer,” i.e., a compound of Formula 3S-III:

In another such embodiment, a compound of Formula III is adiastereomeric mixture having R-stereochemistry at the 3-position,referred to as the “3R-isomer,” i.e., a compound of Formula 3R-III:

In another such embodiment, a compound of Formula III is in the 2R,3S-isomeric form, i.e., a compound of Formula 2R,3S-III:

In another such embodiment, a compound of Formula III is in the 2S,3S-isomeric form, i.e., a compound of Formula 2S,3S-III:

In another such embodiment, a compound of Formula III is in the2R,3R-isomeric form, i.e., a compound of Formula 2R,3R-III:

In another such embodiment, a compound of Formula III is in the2S,3R-isomeric form, i.e., a compound of Formula 2R,3R-III.

In one embodiment, particularly as it relates to a compound of FormulaIII, or a stereoisomer or mixture of stereoisomers thereof, RX is loweralkyl halide. In another such embodiment, RX is a lower alkyl iodide. Instill another such embodiment, RX is methyl iodide. In certainembodiments, about a 3-fold excess or more of RX is used in the reactionbetween a compound of Formula IV and RX, e.g., about a 4-fold, about a5-fold, about a 6-fold, about a 7-fold, about a 8-fold, about a 9-foldor about a 10-fold excess or more.

In additional embodiments, the reaction is carried out in the presenceof a base, which may be, for example, potassium carbonate.

In additional embodiments, the reaction is carried out in a solventselected from the group consisting of C₁-C₄ alcohol (e.g., methanol),tetrahydrofuran, acetonitrile and dichloromethane.

In additional embodiments, the reaction is carried out a temperaturefrom about 20° C. to about 50° C., such as a temperature from about 37°C. to about 42° C.

In additional embodiments, the reaction mixture is filtered and thesolvent is removed by evaporation to give a compound of Formula III.

In another embodiment, R¹ is selected from the group consisting ofhydrogen, optionally substituted lower alkyl, aralkyl, and optionallysubstituted aryl. In another such embodiment, R¹ is selected from thegroup consisting of lower alkyl, aralkyl, and optionally substitutedaryl, including but not limited to, methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl, andparticularly isobutyl.

In additional embodiments, R² and R³ are independently selected from thegroup consisting of lower alkyl, aralkyl, and aryl, including but notlimited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl, and particularlymethyl.

The progress of the reaction between a compound of Formula IV, orstereoisomer or mixture of stereoisomers thereof, and RX can bemonitored by analytical methods known in the art such as TLC, LC, LC/MS,HPLC, NMR, etc., according to routine protocols that will be familiar tothose of ordinary skill in the art. A compound of Formula III, or astereoisomer or mixture of stereoisomers thereof can be isolated andpurified by any means known in the art such normal- and reverse-phasecolumn chromatography (e.g., column chromatography on silica gel orreverse-phase HPLC), crystallization, extraction, etc., according toroutine protocols that will be familiar to those of ordinary skill inthe art. The product thus isolated can be subjected to furtherpurification (e.g., recrystallization) until the desired level of purityis achieved. Thus, in certain embodiments, the compounds of Formula III,or the stereoisomers or mixture of stereoisomers thereof, have a purityof about 90% or more, e.g., about 91%, about 92%, about 93%, about 94%,about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% ormore. In such embodiments, the level of purity can be determined by anysuitable analytical technique, e.g., NMR, LC, HPLC, etc., according toprotocols that will be familiar to the ordinarily skilled artisan.

In additional embodiments, the present invention also provides methodsfor separating the stereoisomers, e.g., diastereomers, of a compound ofFormula III. Thus, in certain such embodiments, the present inventionprovides methods for the preparation of a compound of Formula 2S,3S-IIIfrom a compound of Formula 3S-III, for example, comprising isolating acompound of Formula 2S,3S-III, i.e., the 2S,3S-isomer, that issubstantially free from a compound of Formula 2R,3S-III, i.e., the2R,3S-isomer. In additional embodiments, the invention provides methodsfor the preparation of a compound of Formula 2R,3S-III from a compoundof Formula 3S-III, for example, comprising isolating a compound ofFormula 2R,3S-III that is substantially free from a compound of Formula2S,3S-III. In additional embodiments, the invention provides methods forthe preparation of a compound of Formula 2R,3R-III from a compound ofFormula 3R-III, for example, comprising isolating a compound of Formula2R,3R-III, i.e., the 2R,3R-isomer, that is substantially free from acompound of Formula 2S,3R-III, i.e., the 2S,3R-isomer. In additionalembodiments, the invention provides methods for the preparation of acompound of Formula 2S,3R-III from a compound of Formula 3R-III, forexample, comprising isolating a compound of Formula 2S,3R-III that issubstantially free from a compound of Formula 2R,3R-III.

According to this aspect of the invention, the diastereomers of acompound of Formulae 3S-III or 3R-III can be isolated by any suitableseparation technique known in the art of chemical synthesis, e.g.,chromatography or crystallization, as described in further detail hereinbelow. In one such embodiment, the diastereomers are isolated bychromatography, e.g., reverse phase chromatography. In another suchembodiment, the diastereomers are isolated by crystallization.

In one embodiment, a compound of Formula 2R,3S-III is isolated with adiastereomeric excess of about 90% or more, e.g., about 91%, about 92%,about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about99% or more. In another embodiment, a compound of Formula 2S,3S-III isisolated with a diastereomeric excess of about 90% or more, e.g., about91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%,about 98%, about 99% or more. In still another embodiment, a compound ofFormula 2R,3R-III is isolated with a diastereomeric excess of about 90%or more, e.g., about 91%, about 92%, about 93%, about 94%, about 95%,about 96%, about 97%, about 98%, about 99% or more. In still anotherembodiment, a compound of Formula 2S,3R-III is isolated with adiastereomeric excess of about 90% or more, e.g., about 91%, about 92%,about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about99% or more. In such embodiments, the level of diastereomeric excess canbe determined by any suitable analytical technique, e.g., NMR, LC, HPLC,etc., according to protocols that will be familiar to the ordinarilyskilled artisan.

In one particular embodiment according to this aspect of the invention,the diastereomers of a compound of Formulae 3S-III or 3R-III areseparated by crystallization. In one such embodiment, thecrystallization comprises:

-   (a) dissolving a compound of Formulae 3S-III or 3R-III in a solvent    or solvent system, i.e., a mixture of solvents, e.g.,    methanol/water, ethanol/water, tetrahydrofuran/water,    acetonitrile/water, etc., to give a solution;-   (b) allowing precipitation to occur thereby forming a precipitate;    and-   (c) separating crystalline product in said precipitate from the    solution.

In certain such embodiments, the solution is a homogeneous solution,i.e., the compound of Formulae 3S-III or 3R-III is completely dissolved.In certain embodiments, the solvent or solvent system is selected fromthe group consisting of dichloromethane, methanol, methanol/water,ethanol, ethanol/water, isopropanol, isopropanol/water, tetrahydrofuran,tetrahydrofuran/water, acetonitrile and acetonitrile/water.

In other embodiments of the invention, precipitation of the desiredproduct, i.e., a compound of Formulae 2R,3S-III, 2S,3S-III, 3R,3R-III or2S,3R-III, is induced by adding an anti-solvent to said solution. Incertain such embodiments, the anti-solvent is selected from the groupconsisting of hexane, ethyl acetate, acetone, methyl ethyl ketone, andmethyl t-butyl ether, and particularly ethyl acetate.

In additional embodiments, precipitation of the product duringcrystallization is induced by cooling the solution, for example to about10° C., to about 5° C., or to about 0° C. In certain such embodiments,the solution is heated before, during, or after the addition of theanti-solvent.

In certain additional embodiments, the crystalline product is isolatedby filtration.

The present invention also provides methods for the preparation of acompound of Formula IV, the methods comprising:

-   (a) removing R⁶, or removing R⁶ and R⁷, from a compound of Formula V

particularly wherein:

-   R⁶ is an amine protecting group; and-   R⁷ is selected from the group consisting of hydrogen and an amine    protecting group, or R⁶ and R⁷ taken together represent an amine    protecting group; and R¹, R^(2a) and R^(3a) have the meanings as    described above for Formula III, to give a compound of Formula IV,    or stereoisomer or mixture of stereoisomers thereof; and-   (b) isolating said compound of Formula IV; or-   (C) using said compound of Formula IV in the next reaction without    isolation.

In one such embodiment, a compound of Formula V is a diastereomericmixture having S-stereochemistry at the 3-position, i.e. a compound ofFormula 3S-V:

In another such embodiment, a compound of Formula V is a diastereomericmixture having R-stereochemistry at the 3-position, i.e. a compound ofFormula 3R-V:

As it relates to a compound of Formula V, or a stereoisomer or mixtureof stereoisomers thereof, in certain embodiments, —NR⁶R⁷ represents amono-protected amine, i.e., R⁶ is an amine protecting group, and R⁷ ishydrogen. In one such embodiment, R⁶ is selected from the groupconsisting of carbobenzyloxy and tert-butyloxycarbonyl, i.e., —NR⁶R⁷ is—N(H)CBz or —N(H)Boc. In another such embodiment, —NR⁶R⁷ represents adi-protected amine wherein R⁶ and R⁷ are each independently an amineprotecting group. In still another such embodiment, R⁶ is selected fromthe group consisting of carbobenzyloxy and tert-butyloxycarbonyl, and R⁷is benzyl, i.e., —NR⁶R⁷ is —N(Cbz)Bn or —N(Boc)Bn. In additionalembodiments, —NR⁶R⁷ represents a di-protected amine wherein R⁶ and R⁷taken together represent an amine protecting group such as a phthalimidegroup.

In one particular embodiment, R⁶ and R⁷ are benzyl, i.e., —NR⁶R⁷ is—NBn₂.

In one such embodiment, the benzyl groups are removed, i.e., the amineis deprotected to give —NH₂, for example, under an atmosphere ofhydrogen gas using palladium on carbon as the catalyst. In such oneembodiment the deprotection is carried out in a solvent selected fromthe group consisting of C₁-C₄ alcohol (e.g., methanol), tetrahydrofuran,acetonitrile and dichloromethane. In certain such embodiments, thedeprotection is carried out a temperature from about 20° C. to about 50°C., e.g., about 37° C. to about 42° C.

In certain such embodiments, the reaction mixture is filtered through apad of celite and the solvent(s) are removed by evaporation. In otherembodiments, however, a compound of Formula IV is used in the nextsynthetic step without additional purification.

In certain embodiments according to this aspect of the invention, R¹ ina compound of Formula V, or a stereoisomer or mixture or stereoisomersthereof, is selected from the group consisting of hydrogen, optionallysubstituted lower alkyl, aralkyl, and optionally substituted aryl. Inother embodiments, R¹ is selected from the group consisting of loweralkyl, aralkyl, and optionally substituted aryl. In certain suchembodiments, R¹ is selected from the group consisting of methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl,and phenyl, and particularly isobutyl. In additional embodiments, R² andR³ are independently selected from the group consisting of lower alkyl,aralkyl, and aryl, including but not limited to, methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, isopenyl,benzyl, and phenyl. In certain such embodiments, R² and R³ are methyl.

The progress of the removal, i.e., deprotection, of the R⁶ group, or R⁶and R⁷ groups, from a compound of Formula V to give a compound ofFormula IV can be monitored by analytical methods known in the art suchas TLC, LC, LC/MS, HPLC, NMR, etc., according to routine protocols thatwill be familiar to those of ordinary skill in the art. A compound ofFormula IV, or a stereoisomer or mixture of stereoisomers thereof, canbe isolated and purified by any means known in the art such normal- andreverse-phase column chromatography (e.g., column chromatography onsilica gel or reverse-phase HPLC), crystallization, extraction, etc.,according to routine protocols that will be familiar to those ofordinary skill in the art. The product thus isolated can be optionallysubjected to further purification (e.g., recrystallization) until thedesired level of purity is achieved. Thus, in certain embodiments of theinvention, a compound of Formula IV, or a stereoisomer or mixture ofstereoisomers thereof, has a purity of about 90%, about 91%, about 92%,about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about99%, about 99.5% or more. In such embodiments, the level of purity canbe determined by any suitable analytical technique, e.g., NMR, LC, HPLC,etc., according to protocols that will be familiar to the ordinarilyskilled artisan.

The present invention also provides methods for the preparation ofβ-hydroxy-γ-aminophosphonates of Formula V, the methods comprising:

-   (a) reducing a β-keto-γ-aminophosphonate of Formula VI:

particularly wherein R¹, R^(2a) and R^(3a) have the meanings asdescribed above for Formula III, and R⁶ and R⁷ have the meanings asdescribed above for Formula V, to give a compound of Formula V, or astereoisomer or mixture of stereoisomers thereof; and

-   (b) isolating said compound of Formula V; or-   (c) using said compound of Formula V in the next reaction without    isolation.

In one such embodiment, a compound of Formula VI has S-stereochemistryat the 3-position, i.e. a compound of Formula 3S-VI:

In another such embodiment, a compound of Formula VI hasR-stereochemistry at the 3-position, i.e. a compound of Formula 3R-VI:

As it relates to a compound of Formula VI, or a stereoisomer or mixtureof stereoisomers thereof, in certain embodiments, the oxo group, i.e.,C═O, of a compound of Formula VI is reduced using a borohydride reducingagent. In one such embodiment, the borohydride reducing agent is sodiumborohydride.

In additional embodiments, the reaction is carried out in a solventsystem comprising a C₁-C₄ alcohol (e.g., methanol) and tetrahydrofuran.In one such embodiment, the solvent system comprises about 5% to about15% tetrahydrofuran in methanol (v/v). In another such embodiment, thesolvent system comprises about 10% tetrahydrofuran and about 90%methanol (v/v).

In additional embodiments, the reduction is carried out a temperaturefrom about −30° C. to about 20° C., and particularly from about −10° C.to about 5° C.

In certain embodiments according to this aspect of the invention, thecompound of Formula V is used in the next reaction without purification.

In certain embodiments according to this aspect of the invention, R¹ isselected from the group consisting of hydrogen, optionally substitutedlower alkyl, aralkyl, and optionally substituted aryl. In one suchembodiment, R¹ is selected from the group consisting of lower alkyl,aralkyl and optionally substituted aryl. In certain such embodiments, R¹is selected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,and particularly isobutyl.

In additional embodiments, R² and R³ are independently selected from thegroup consisting of lower alkyl, aralkyl, and aryl. In one suchembodiment, R² and R³ are independently selected from the groupconsisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopenyl, benzyl, and phenyl. In another suchembodiment, R² and R³ are methyl. In still another such embodiment, R⁶and R⁷ are benzyl.

The progress of the reduction of a β-keto-γ-aminophosphonate of FormulaVI to a β-hydroxy-γ-aminophosphonate of Formula V can be monitored byanalytical methods known in the art such as TLC, LC, LC/MS, HPLC, NMR,etc., according to routine protocols that will be familiar to those ofordinary skill in the art. A compound of Formula V, or a stereoisomer ormixture of stereoisomers thereof, can be isolated and purified by anymeans known in the art such normal- and reverse-phase columnchromatography (e.g., column chromatography on silica gel orreverse-phase HPLC), crystallization, extraction, etc., according toroutine protocols that will be familiar to those of ordinary skill inthe art. The product thus isolated can be optionally subjected tofurther purification (e.g., recrystallization) until the desired levelof purity is achieved. Thus, in certain embodiments of the invention, acompound of Formula V has a purity of about 90% or more, e.g., about91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%,about 98%, about 99% or more. In such embodiments, the level of puritycan be determined by any suitable analytical technique, e.g., NMR, LC,HPLC, etc., according to protocols that will be familiar to theordinarily skilled artisan.

Alternatively, a compound of Formula V can be used in a subsequentchemical transformation without further purification.

The present invention also provides methods for the preparation of acompound of Formula VI, the methods comprising:

-   (a) condensing of a compound of Formula VII:

with a compound of Formula VIII:

particularly wherein R¹, R^(2a) and R^(3a) have the meanings asdescribed above for Formula III, R⁶ and R⁷ have the meanings asdescribed above for Formula V, and R⁸ is selected from the groupconsisting of optionally substituted alkyl, aralkyl, and optionallysubstituted aryl, to give a compound of Formula VI, or a stereoisomer ormixture of stereoisomers thereof; and

-   (b) isolating said compound of Formula VI; or-   (c) using said compound of Formula VI in the next reaction without    isolation.

In one such embodiment, a compound of Formula VII is the S-isomer, i.e.a compound of Formula S-VII:

In another such embodiment, a compound of Formula VII is the R-isomer,i.e. a compound of Formula R-VII:

As it relates to a compound of Formula VII, or a stereoisomer or mixtureof stereoisomers thereof, in certain embodiments, the condensation witha compound of Formula VIII is carried out in an inert organic solvent.In one such embodiment, the inert organic solvent is tetrahydrofuran.

In additional embodiments, the condensation is carried out attemperature ranging from about −78° C. to about −20° C., particularly atabout −50° C.

In certain embodiments according to this aspect of the invention, R¹ isselected from the group consisting of hydrogen, optionally substitutedlower alkyl, aralkyl, and optionally substituted aryl. In another suchembodiment, R¹ is selected from the group consisting of lower alkyl,aralkyl, and optionally substituted aryl. In still another suchembodiment, R¹ is selected from the group consisting of methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl,and phenyl, and particularly isobutyl.

In additional embodiments, R² and R³ are independently selected from thegroup consisting of lower alkyl, aralkyl, and aryl. In another suchembodiment, R² and R³ are independently selected from the groupconsisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, isopentyl, benzyl, and phenyl. In still anothersuch embodiment, R² and R³ are methyl.

In additional embodiments, R⁶, R⁷ and R⁸ are benzyl.

The progress of the condensation of a compound of Formula VII with acompound of Formula VIII can be monitored by analytical methods known inthe art such as TLC, LC, LC/MS, HPLC, NMR, etc., according to routineprotocols that will be familiar to those of ordinary skill in the art. Acompound of Formula VI, or a stereoisomer or mixture of stereoisomersthereof, can be isolated and purified by any means known in the art suchnormal- and reverse-phase column chromatography (e.g., columnchromatography on silica gel or reverse-phase HPLC), crystallization,extraction, etc., according to routine protocols that will be familiarto those of ordinary skill in the art. The product thus isolated can beoptionally subjected to further purification (e.g., recrystallization)until the desired level of purity is achieved. Thus, in certainembodiments of the invention, a compound of Formula VI, or astereoisomer or mixture of stereoisomers thereof, has a purity of about90% or more, e.g., about 91%, about 92%, about 93%, about 94%, about95%, about 96%, about 97%, about 98%, about 99 or more. In suchembodiments, the level of purity can be determined by any suitableanalytical technique, e.g., NMR, LC, HPLC, etc., according to protocolsthat will be familiar to the ordinarily skilled artisan.

Alternatively, a compound of Formula VI can be used in a subsequentchemical transformation without further purification.

The present invention also provides methods for the preparation of acompound of Formula VIII, the methods comprising:

-   (a) condensing a compound of Formula IX:

with LiR⁹, particularly wherein R^(2a) and R^(3a) have the meanings asdescribed above for Formula III and R⁹ is selected from the groupconsisting of lower alkyl and aryl, to give a compound of Formula VIII;and

-   (b) isolating said compound of Formula VIII; or-   (c) using said compound of Formula VIII without isolation.

In one such embodiment according to this aspect of the invention, R^(2a)and R^(3a) are independently selected from the group consisting of loweralkyl, and aralkyl. In another such embodiment, R^(2a) and R^(3a) arelower alkyl, particularly methyl. In one embodiment, R⁹ is lower alkyl,particularly n-butyl (i.e., in that particular embodiment, LiR⁹ isn-butyl lithium).

In certain embodiments, the condensation is carried out in an inertorganic solvent such as tetrahydrofuran.

In certain embodiments, the condensation is carried out at temperatureranging from about −78° C. to about −20° C., more particularly at about−60° C. to about −30° C., still more particularly at about −60° C. toabout −50° C., and still more particularly at about −50° C.

The progress of the condensation of a compound of Formula IX with LiR⁹can be monitored by analytical methods known in the art such as TLC, LC,LC/MS, HPLC, NMR, etc., according to routine protocols that will befamiliar to those of ordinary skill in the art. A compound of FormulaVIII is typically used in situ without isolation or purification.

The present invention also provides methods for the preparation of acompound of Formula X:

particularly wherein R and R¹ have the meanings as described above forFormula III, R^(2b) and R^(3b) are selected from the group consisting ofhydrogen and monovalent pharmaceutically acceptable cation; or takentogether R^(2b) and R^(3b) represent a divalent pharmaceuticallyacceptable cation; or X⁻ and R^(2b) are absent (i.e., a compound ofFormula X is a zwitterion), the method comprising:

-   (a) removing R^(2a) and R^(3a) from a compound of Formula III, to    give a compound of Formula X, or a stereoisomer or mixture of    stereoisomers thereof; and-   (b) isolating said compound of Formula X.

In one such embodiment, a compound of Formula X is a diastereomericmixture having S-stereochemistry at the 3-position, i.e., a compound ofFormula 3S-X:

In another such embodiment, a compound of Formula X is a diastereomericmixture having R-stereochemistry at the 3-position, i.e., a compound ofFormula 3R-X:

In another such embodiment, a compound of Formula X is the 2R,3S-isomer, i.e., a compound of Formula 2R,3S-X:

In another such embodiment, a compound of Formula X is the 2S,3S-isomer, i.e., a compound of Formula 2S,3S-X:

In another such embodiment, a compound of Formula X is the 2R,3R-isomer, i.e., a compound of Formula 2R,3R-X:

In another such embodiment, a compound of Formula X is the 2S,3R-isomer, i.e., a compound of Formula 2S,3R-X:

As it relates to a compound of Formula X, or a stereoisomer or mixtureof stereoisomers thereof, in certain embodiments, R is lower alkyl,particularly methyl. In additional embodiments, R¹ is selected from thegroup consisting of hydrogen, optionally substituted lower alkyl,aralkyl, and optionally substituted aryl. In another such embodiment, R¹is selected from the group consisting of lower alkyl, aralkyl, andoptionally substituted aryl, particularly methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl.In still another such embodiment, R¹ is isobutyl. In additionalembodiments, R^(2b) and R^(3b) are hydrogen.

In certain embodiments according to this aspect of the invention, R^(2b)and R^(3b) are each a monovalent pharmaceutically acceptable cation,including but not limited to Na⁺ and K⁺.

In another embodiment, R^(2b) and R^(3b) taken together represent adivalent pharmaceutically acceptable cation, including but not limitedto Mg²⁺ and Ca²⁺.

In another embodiment, X⁻ and R^(2b) are absent and the compound ofFormula X, or stereoisomer or mixture of stereoisomers thereof, is azwitterion.

In additional embodiments, R^(2a) and R^(3a) of a compound of FormulaIII, or a stereoisomer or mixture of stereoisomers thereof, are benzyl.In certain embodiments, the benzyl groups are removed under anatmosphere of hydrogen gas, and palladium on carbon is the catalyst. Inone embodiment the benzyl groups are removed in a solvent selected fromthe group consisting of C₁-C₄ alcohol (e.g., methanol), tetrahydrofuran,acetonitrile and dichloromethane.

In additional embodiments, the benzyl groups are removed at temperaturefrom about 20° C. to about 50° C., e.g., from about 37° C. to about 42°C.

In additional embodiments, the reaction mixture is filtered through apad of celite and the solvent(s) are removed by evaporation to give acompound of Formula X, or a stereoisomer or mixture of stereoisomersthereof wherein R^(2b) and R^(3b) are hydrogen.

In additional embodiments, R^(2a) and R^(3a) of a compound of FormulaIII, or a stereoisomer or mixture of stereoisomers thereof, are methyl.In certain embodiments, the methyl groups are removed using bromotrimethylsilane. In one embodiment the methyl groups are removed in asolvent selected from the group consisting of tetrahydrofuran anddichloromethane.

In additional embodiments, the methyl groups are removed at temperaturefrom about 20° C. to about 50° C., e.g., from about 37° C. to about 42°C.

In additional embodiments, the solvents are removed by evaporation. Inadditional embodiments, the reaction mixture is dissolved in water.

In additional embodiments, the reaction mixture is filtered and thesolvent(s) are removed by evaporation to give a compound of Formula X,or a stereoisomer or mixture of stereoisomers thereof, wherein R^(2b)and R^(3b) are hydrogen.

The progress the removal of the R^(2a) and R^(3a) groups from a compoundof Formula III to give a compound of Formula X can be monitored byanalytical methods known in the art such as TLC, LC, LC/MS, HPLC, NMR,etc., according to routine protocols that will be familiar to those ofordinary skill in the art. A compound of Formula X, or a stereoisomer ormixture of stereoisomers thereof, can be isolated and purified by anymeans known in the art such normal- and reverse-phase columnchromatography (e.g., column chromatography on silica gel orreverse-phase HPLC), crystallization, extraction, etc., according toroutine protocols that will be familiar to those of ordinary skill inthe art. The product thus isolated can be optionally subjected tofurther purification (e.g., recrystallization) until the desired levelof purity is achieved. Thus, in certain embodiments of the invention, acompound of Formula X, or stereoisomer or mixture of stereoisomersthereof, has a purity of about 90%, about 91%, about 92%, about 93%,about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, about99.5% or more. In such embodiments, the level of purity can bedetermined by any suitable analytical technique, e.g., NMR, LC, HPLC,etc., according to protocols that will be familiar to the ordinarilyskilled artisan.

In additional embodiments, the invention provides methods for thepreparation of a compound of Formula VII, the method comprising:

-   (a) protecting the amine of an amino acid of Formula XI; and-   (b) esterifying the carboxylic acid of an amino acid of Formula XI:

to give a compound of Formula VII, particularly wherein R¹ has themeaning as described above for Formula III; and

-   (c) isolating said compound of Formula VII; or-   (d) using said compound of Formula VII in the next reaction without    isolation.

In one such embodiment, the compound of Formula XI is the S-isomerhaving Formula S-XI:

In another such embodiment, the compound of Formula XI is the R-isomerhaving Formula R-XI:

In certain embodiments, the protection and esterification of a compoundof Formula XI are accomplished simultaneously by condensation with abenzyl halide. In one such embodiment, about a 3-fold excess or more(i.e., about a 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-foldexcess or more) of a benzyl halide, such as benzyl bromide, is used inthe condensation. In certain embodiments, the condensation with a benzylhalide is carried out in a solvent system selected from the groupconsisting of a C₁-C₄-alcohol/water or acetonitrile/water. In one suchembodiment, the C₁-C₄-alcohol is methanol. In a further embodiment, thesolvent system comprises about 5% to about 50% solvent in water. Inadditional embodiments, the condensation with a benzyl halide is carriedout in the presence of a base, such as potassium carbonate.

In certain aspects of the present invention, compounds of Formula I andII, or stereoisomers or mixtures of stereoisomers thereof, inhibitcarnitine acyltransferases. Therefore, it is contemplated by the presentinvention that such compounds will be useful therapeutic agents, e.g.,anticholesteremic, hypolipidemic, antidiabetic or antiobesity agents.Thus, the present invention provides compounds, compositions and methodsfor use in patients, particularly mammals such as humans, havingdiseases, conditions or disorders linked with disorders in fatty acidmetabolism.

Thus, in certain embodiments of the invention, the compound of Formula Iis selected from the group consisting of:

or pharmaceutically acceptable salts, esters, amides or zwitterions orother pharmaceutically acceptable derivatives, variants or formsthereof.

In additional embodiments, the compounds, compositions and methods ofthe present invention are used in methods provided by the invention totreat diseased cells, tissues, organs, organ systems, or pathologicalconditions, disorders, and/or disease states in patients (e.g., amammalian subject including, but not limited to, humans and veterinaryanimals). In this regard, various pathological conditions and/or diseasestates are amenable to treatment, amelioration or prevention using themethods and compositions provided by the present invention.

In certain embodiments, the pathological condition and/or disease statethat is amenable to treatment, amelioration or prevention using thepresent methods and compositions is one that is linked with disorders infatty acid metabolism. In other embodiments, the pathological conditionsand/or diseases state that are amenable to treatment, amelioration orprevention using the present methods and compositions includes, but isnot limited to, hyperlipoproteinemia, hyperlipidemia, cardiac disorders,e.g., myocardial dysfunction, renal anemia, Alzheimer's disease,non-insulin dependent diabetes mellitus and obesity. Methods accordingto such aspects of the invention include, for example, (a) identifying apatient suffering from, afflicted with, or predisposed to a condition,disorder or disease state associated with disorders in fatty acidmetabolism (including but not limited to the conditions, disorders anddisease states noted above); and (b) administering to said patient oneor more compounds or pharmaceutical compositions of the invention in anamount effective to treat, ameliorate and/or prevent the appearance,effects, symptoms and/or progression of the disease, condition ordisorder in the patient. According to certain such aspects of theinvention, the approximate dosage form(s), mode(s) of administration,dosage amounts and dosing regimen(s) for use in these methods includethose described herein, although additional suitable dosage forms, modesof administration, dosage amounts and dosing regimens will be familiarto those of ordinary skill in the relevant arts and/or can beempirically determined by the clinical practitioner using routinemethods known to the ordinarily skilled artisan based on the guidanceprovided herein and in view of the information that is readily availablein the art.

Compositions within the scope of the present invention include allcompositions wherein one or more of the compounds of the presentinvention are contained in an amount which is effective to achieve itsintended purpose. While individual needs vary, determination of optimalranges of effective amounts of each component is within the expertise ofthose of ordinary skill in the art. Typically, the compounds may beadministered to mammals, e.g. humans, orally at a dose of about 0.0025to about 50 mg/kg, or an equivalent amount of the pharmaceuticallyacceptable salt or ester thereof. For example, about 0.01 to about 25mg/kg can be orally administered to treat, ameliorate, or prevent suchdisorders. For intramuscular injection, the dose is generally aboutone-half of the oral dose, for example, a suitable intramuscular dosewould be about 0.0025 to about 25 mg/kg, e.g., from about 0.01 to about5 mg/kg.

Compositions with the scope of the present invention also include allcompositions wherein one or more of the compounds of the presentinvention are combined with one or more additional therapeutic agents(e.g., anticholesterolemics, anticoagulants, anti-obesity oranti-diabetic drugs) in therapeutically effective amounts. In additionto active agents (e.g., one or more compounds of the invention and oneor more additional therapeutic agents), such compositions can optionallycomprise one or more pharmaceutical excipients well-known in therelevant arts. Typically, such compositions are administered orally. Theoptimal amounts of each active agent in the composition can bedetermined by the clinical practioner using routine methods known to theordinarily skilled artisan based on the guidance provided herein and inview of the information that is readily available in the art.

The unit oral dose may comprise from about 0.01 to about 1000 mg, e.g.,about 0.1 to about 100 mg of the compound. The unit dose may beadministered one or more times daily as one or more tablets or capsuleseach containing from about 0.1 to about 10, conveniently about 0.25 to50 mg of the compound or its solvates.

In a topical formulation, the compound may be present at a concentrationof about 0.01 to 100 mg per gram of carrier. In one embodiment, thecompound is present at a concentration of about 0.07-1.0 mg/ml, e.g.,about 0.1-0.5 mg/ml, e.g., about 0.4 mg/ml.

In addition to administering the compound as a raw chemical, thecompounds of the invention may be administered as part of apharmaceutical composition comprising one or more compounds of theinvention and one or more suitable pharmaceutically acceptable carriers,such as one or more excipients or auxiliaries which facilitateprocessing of the compounds into preparations which can be usedpharmaceutically. Preferably, such pharmaceutical compositions containfrom about 0.01 to 99 percent, e.g., from about 0.25 to 75 percent ofactive compound(s), together with the excipient(s), particularly thosecompositions which can be administered orally or topically and which canbe used for the preferred type of administration, such as tablets,dragees, slow release lozenges and capsules, mouth rinses and mouthwashes, gels, liquid suspensions, and also preparations which can beadministered rectally, such as suppositories, as well as suitablesolutions for administration by parenteral administration, e.g., viaintravenous infusion, intramuscular or subcutaneous injection.

The pharmaceutical compositions of the invention may be administered toany patient who may experience the beneficial effects of the compoundsand/or compositions of the invention. Foremost among such patients arehumans, although the invention is not intended to be so limited. Otherpatients include veterinary animals (cows, sheep, pigs, horses, dogs,cats and the like).

The compounds and pharmaceutical compositions of the invention may beadministered by any means that achieve their intended purpose. Forexample, administration may be by parenteral, subcutaneous, intravenous,intramuscular, intradermal, intraperitoneal, transdermal, buccal,sublingual, intrathecal, intracranial, intranasal, ocular, pulmonary(e.g., via inhalation) or topical routes. Alternatively, orconcurrently, administration may be by the oral route. The dosageadministered will be dependent upon the age, health, and weight of therecipient, kind of concurrent treatment, if any, frequency of treatment,and the nature of the effect desired.

Suitable oral pharmaceutical compositions of the present invention aremanufactured in a manner which is itself well-known in the art, forexample, by means of conventional mixing, granulating, dragee-making,dissolving, or lyophilizing processes. Thus, solid pharmaceuticalpreparations for oral use can be obtained by combining one or more ofthe compounds of the invention and optionally one or more additionalactive pharmaceutical ingredients with one or more solid excipients,optionally grinding the resulting mixture and processing the mixture ofgranules, after adding suitable auxiliaries, if desired or necessary, toobtain tablets or dragee cores.

Suitable excipients are, in particular, fillers such as saccharides, forexample lactose, sucrose, fructose and the like; sugar alcohols such asmannitol, sorbitol, or xylitol and the like; cellulose preparationsand/or calcium phosphates, for example tricalcium phosphate or calciumhydrogen phosphate; as well as binders such as starch paste, using, forexample, maize starch, wheat starch, rice starch, potato starch,gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose,sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired,disintegrating agents may be added such as the above-mentioned starchesand also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar,or alginic acid or a salt thereof, such as sodium alginate. Auxiliariesare, above all, flow-regulating agents and lubricants, for example,silica, talc, stearic acid or salts thereof, such as magnesium stearateor calcium stearate, and/or poly(ethylene glycol). Dragee cores areprovided with suitable coatings which, if desired, are resistant togastric juices. For this purpose, concentrated saccharide solutions maybe used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, poly(ethylene glycol) and/or titanium dioxide, lacquersolutions and suitable organic solvents or solvent mixtures. In order toproduce coatings resistant to gastric juices, solutions of suitablecellulose preparations such as acetylcellulose phthalate orhydroxypropylmethyl-cellulose phthalate, can be used. Dye stuffs orpigments may be added to the tablets or dragee coatings, for example,for identification or in order to characterize combinations of activeingredients or doses thereof.

Other pharmaceutical preparations which can be used orally includepush-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer such as glycerol or sorbitol. In certainembodiments, the push-fit capsules can comprise one or more of thecompounds of the invention in the form of granules which may be mixedwith fillers such as lactose, binders such as starches, and/orlubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, one or more pharmaceutical ingredients(e.g., one or more compounds of the invention and optionally one or moreadditional active pharmaceutical ingredients) are preferably dissolvedor suspended in suitable liquids, such as fatty oils, or liquidparaffin. In addition, stabilizers may be added.

Suitable pharmaceutical preparations which can be used rectally include,for example, suppositories, which comprise a combination of one or moreof the compounds of the invention with a suppository base. Suitablesuppository bases are, for example, natural or synthetic triglycerides,or paraffin hydrocarbons. In addition, it is also possible to usegelatin rectal capsules which consist of a combination of the activecompounds with a base. Possible base materials include, for example,liquid triglycerides, poly(ethylene glycols), or paraffin hydrocarbons.

In addition to the solid dosage forms disclosed throughout, the presentinvention also provides chewable oral formulations. In certain suchembodiments, the formulations will comprise (or consist essentially of)an effective amount of one or more compounds of the invention along withsuitable excipients that allow the formulations to be chewed by thepatient. In additional embodiments, the formulations can furthercomprise one or more taste-masking or sweetening agents, such as thosedescribed herein. In one embodiment, sucralose is used in the chewableformulations. Additional active agents, such as those described herein,can also optionally be added to the chewable formulations. The amount ofone or more compounds of the invention, other optional active agents(e.g., anticholesterolemics, anticoagulants, anti-obesity oranti-diabetic drugs), and sweetening agents (e.g., sucralose) in thechewable formulations of the present invention are readily determinableby those of ordinary skill in the art, and include those amounts andcombinations described herein. For example, the chewable formulations ofthe present invention comprise (or consist essentially of) about 0.05%to about 5% of a one or more compounds of the invention, optionallyabout 0.01% to about 10% other active agent(s) (or more as required),and about 0.05% to about 0.15% sucralose. Such chewable formulations areespecially useful in patient populations where compliance is an issue,such as children, the elderly, and patients who may have difficultyswallowing or using spray/inhalable formulations.

The formulations may also contain colorants to improve the appearance ofthe chewable formulations, especially since an attractive colorationimparted by a colorant may improve patient compliance. The relativeamounts of the colorants selected will vary depending upon theparticular hue of the individual colorants and the resultant colordesired.

Any standard pharmaceutically acceptable excipient can be used in thechewable tablet formulations which provides adequate compression such asdiluents (e.g., mannitol, xylitol, maltitol, lactitol, sorbitol,lactose, sucrose, and compressible sugars such as DiPac® (dextrinizedsucrose), available from Austin Products Inc. (Holmdel, N.J.), binders,disintegrants, splitting or swelling agents (e.g., polyvinylpolypyrrolidone, croscarmellose sodium (e.g., Ac-Di-Sol available fromFMC BioPolymer, Philadelphia, Pa.), starches and derivatives, celluloseand derivatives, microcrystalline celluloses, such as Avicel™ PH 101 orAvicel™ CE-15 (a microcrystalline modified with guar gum), bothavailable from FMC BioPolymer, (Philadelphia, Pa.), lubricating agents(e.g., magnesium stearate), and flow agents (e.g., colloidal silicondioxide, such as Cab-O-Sil M5® available from Cabot Corporation, Kokomo,Ind.).

Suitable amounts of sweetener (e.g., sucralose) used in the chewableformulations, will be familiar to, and can be readily determined by,those skilled in the art. In certain embodiments, the sweetener ispresent in an amount from about 0.05% to about 5.0% (e.g., about 0.05%,about 0.1%, about 0.125%, about 0.15%, about 0.2%, about 0.3%, about0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about1%, about 1.25% about 1.5%, about 1.75%, about 2%, about 2.25%, about2.5%, about 2.75%, about 3%, about 3.25%, about 3.5%, about 3.75%, about4%, about 4.25%, about 4.5%, about 4.75% or about 5%). Those or ordinaryskill in the art will appreciate that the amount of sweetener may varydepending on the strength of the particular sweetener used and thelevels approved by the regulatory authorities for use in pharmaceuticalproducts.

Suitable cyclodextrins for use in the chewable formulations of thepresent invention include α, β, or γ cyclodextrins, or an alkylated orhydroxyalkylated derivatives thereof, such as heptakis(2,6-di-o-methyl)-β-cyclodextrin (DIMEB), randomly methylatedβ-cyclodextrin (RAMEB), and hydroxypropyl β-cyclodextrin (HPβCD). Asuitable cyclodextrin is β-cyclodextrin (available from Cerestar USA,Inc., Hammond, Ind. or from Roquette America, Inc., Keokuk. Iowa underthe trade name Kleptose™). If desired, the complex of the activesubstance with cyclodextrin can be prepared in advance, for example, bymalaxating or granulating the compounds of the invention and anyadditional active substance(s) and the cyclodextrin in the presence ofwater, or by preparing an aqueous solution containing the one or morecompounds of the invention and any additional active substance(s) andthe cyclodextrin in the desired molar ratio. Alternatively, thecompound(s) of the invention and any additional active substance(s) andthe cyclodextrin can be simply mixed with other excipients andadjuvants. The molar ratio of the compound(s) and any additional activesubstance(s) to cyclodextrin is suitably from about 1.0:1.0 to about4.0:1.0.

A typical manufacturing process for making either a single layer orbi-layer chewable tablet generally involves blending of the desiredingredients to form a uniform distribution of the compound(s) of theinvention (and any other active agent(s)), excipients (e.g., colorantsand flavoring agents as well as others). If desired, an inclusioncomplex of the compound(s) of the invention and any other activeagent(s) and cyclodextrin (e.g., β-cyclodextrin) may be formed prior toblending into the mixture by malaxating the compound(s) of the inventionand any other active agent(s) and cyclodextrin in the presence of waterin a planetary mixer for about 20 minutes. The mixture is then dried ina drying oven. After drying, the complex is mixed with anycolor/flavoring blend. The blend is then compressed into a single layeror bi-layer tablet using standard methods well-known to those skilled inthe art (e.g., Kilian T-100 tablet press or Courtoy 292/43 rotarybi-layer press). The colorants and flavoring agents may be added to bothlayers to form a uniform presentation of the tablet. Methods forpreparation of chewable tablets and various components for use in thetablets can be found throughout the detailed description section and theExamples of U.S. Patent Publication No. 2003/0215503, the disclosure ofwhich is incorporated by reference herein for all purposes. Additionalchewable/orally dissolving tablets, and methods for their manufacture,are disclosed in U.S. Patent Publication No. 2004/0265372 and U.S. Pat.No. 6,270,790, the disclosures of each of which are incorporated byreference herein for all purposes.

In another embodiment, the present invention provides orallydisintegrating/orodispersible tablets, such as those disclosed in U.S.Pat. No. 6,723,348, the disclosure of which is incorporated herein byreference in its entirety for all purposes. The orallydisintegrating/orodispersible tablets suitably disintegrate in thebuccal cavity upon contact with saliva forming an easy-to-swallowsuspension. Such tablets comprise (or consist essentially of)compound(s) of the invention, and optionally, one or more additionalactive agents (such as those described herein), in the form of coatedgranules, and a mixture of excipients comprising at least onedisintegrating agent, a soluble diluent agent, a lubricant andoptionally a swelling agent, an antistatic (fluid flow) agent, apermeabilising agent, taste-masking agents/sweeteners, flavoring agentsand colors. In certain such embodiments, thedisintegrating/orodispersible tablets comprise the taste-masking agentsucralose. The amounts of compound(s) of the invention, other optionalactive agents (e.g., anticholesterolemics, anticoagulants, anti-obesityor anti-diabetic drugs), and sweetening agents (e.g., sucralose) in theorally disintegrating tablet formulations of the present invention arereadily determinable by those of ordinary skill in the art, and includethose amounts and combinations described herein. For example, the orallydisintegrating tablet formulations of the present invention compriseabout 0.1% to about 0.15% of compound(s) of the invention, optionallyabout 0.01% to about 10% other active agent(s) (or more as required),and about 0.05% to about 0.15% sucralose.

In suitable embodiments, the particles/granules of compound(s) of theinvention (and any other optional active agents) have a particle sizesuch that about 100% of the particles have an average size of less thanabout 50 μm. In suitable such embodiments, compound(s) of the invention(and any other optional active agents) are present as coated granules.

In one embodiment, the disintegrating tablets according to the inventioncomprise coated granules of compound(s) of the invention (andoptionally, one or more additional active agents), a taste-masking agentsuch as sucralose, and a mixture of excipients, the ratio of the mixtureof excipients to the coated granules suitably is about 0.4:1 to about9:1, more suitable about 1.5:1 to about 5:1, or about 2 to 3 parts byweight, the mixture of excipients suitably comprising: at least onedisintegrating agent, a soluble diluent agent, a lubricant, andoptionally a permeabilising agent, a swelling agent, an antistaticagent, flavoring agents and one or more coloring agents.

In suitable embodiments, the disintegrating agent is selected from thegroup consisting of croscarmellose, available as e.g. Ac-di-sol™,crospovidone available as e.g. Kollidon CL™, sodium starch glycolate andmixtures thereof.

According to one embodiment of the invention, the soluble diluent is apolyol having less than 13 carbon atoms and being either in the form ofa directly compressible product with an average particle size of about100 to 500 μm, or in the form of a powder with an average particle sizeof less than about 100 μm, this polyol suitably being selected from thegroup consisting of mannitol, xylitol, sorbitol and maltitol. Theproportion of disintegrating agent suitably is from about 3 to about 15%by weight, e.g., about 5 to about 15% by weight, and in the case of amixture, each disintegrating agent being present between about 1 andabout 10% by weight, e.g., about 5 to about 10% by weight, and theproportion of soluble diluent agent being about 30 to about 90% byweight, e.g., about 40 to about 60% by weight, based in each case on theweight of the tablet.

Suitable lubricants for use in the disintegrating tablets include, butare not limited to, magnesium stearate, stearic acid, sodium stearylfumarate, micronised polyoxyethyleneglycol (micronised Macrogol 6000),leukine, sodium benzoate and mixtures thereof. The amount of lubricantgenerally is from about 0 to about 3%, e.g., from about 1 to about 2% byweight, based on the weight of the tablet. The lubricant can bedispersed within the mixture of excipients, or according to oneembodiment, sprayed over the outer surface of the tablet. Thus,according to one embodiment of the disintegrating tablets of theinvention, the lubricant is in powder form and is, at least in part,disposed on the surface of the tablets.

The permeabilising agent allows the creation of a hydrophilic networkwhich facilitates the penetration of saliva and hence assists thedisintegration of the tablet. Suitable permeabilising agent include, butare not limited to, silica with a high affinity for aqueous solvents,such as colloidal silica (Aerosil™), precipitated silica (Syloid™ FP244), maltodextrins, β-cyclodextrins and mixtures thereof. The amount ofpermeabilising agent suitably is between about 0 and about 5%, e.g.,from about 0.5 to about 2% by weight, based on the weight of the tablet.

A swelling agent can be incorporated in the mixture of excipients.Suitable swelling agents include, but are not limited to, starch,modified starch or microcrystalline cellulose.

An antistatic agent can also be incorporated as a flow aid. Suitableantistatic agents include, but are not limited to, micronised ornon-micronised talc, fumed silica (Aerosil™ R972), colloidal silica(Aerosil™ 200), precipitated silica (Syloid™ FP 244), and mixturesthereof.

According to one such embodiment of the invention, the granules ofcompound(s) of the invention (and optionally, one or more additionalactive agents such as those described herein) are characterized in thatthe granules are coated and comprise microcrystals of compound(s) of theinvention, sucralose, at least one binder, and optionally a diluentagent, an antistatic agent, and a coloring agent. Furthermore, thegranulation excipients can also include disintegrating agents and/orsurfactants.

Suitable binders include, but are not limited to, cellulosic polymers,such as ethylcellulose, hydroxypropylcellulose and hydroxypropylmethylcellulose, acrylic polymers, such as insoluble acrylateammoniomethacrylate copolymer, polyacrylate or polymethacryliccopolymer, povidones, copovidones, polyvinylalcohols, alginic acid,sodium alginate, starch, pregelatinized starch, sucrose and itsderivatives, guar gum, poly(ethylene glycol), for example an acrylicpolymer, such as Eudragit™ E100, and mixtures thereof.

Optionally, in order to enhance the granulation of the compound(s) ofthe invention (and one or more additional active agents) or one of itspharmaceutically acceptable salts, a diluent agent can be used. Suitablediluent agents include, but are not limited to, microcrystallinecellulose, sucrose, dicalcium phosphate, starches, lactose and polyolsof less than 13 carbon atoms, such as mannitol, xylitol, sorbitol,maltitol, pharmaceutically acceptable amino acids, such as glycin, andtheir mixtures.

In one embodiment, a granule of compound(s) of the invention (as well asany additional active agents, such as those described herein), can be inthe form of a core of granulated microcrystals of compound(s) of theinvention, coated with at least one layer comprising compound(s) of theinvention. Such a coated core is characterized in that the core and thelayer comprise each from 70% to 95%, preferably 80% to 95% by weight ofcompound(s) of the invention, the balance to 100% being formed with atleast one binder and optionally sucralose, and that the coated core issuitably a sphere. See e.g., French patent application FR 00 14803, thedisclosure of which is incorporated by reference herein.

In one embodiment of the invention, the granules can comprise (orconsist essentially of): from about 10% to about 95%, e.g., from about50% to about 70% of one or more compounds of the invention andoptionally one or more additional active agents, such as those describedherein, at most about 20% by weight of the binder, relative to theweight of the one or more compounds of the invention, at most about 5%,suitably about 2% by weight of the antistatic agent, relative to theweight of said granules, suitably about 0.05% to about 5% sucralose andoptionally a diluent agent for the balance to 100%.

The granules can also be coated with a coating composition comprising atleast one coating polymer selected from the group consisting ofcellulosic polymers, acrylic polymers and their mixtures. Among thecellulosic polymers, ethylcellulose, hydroxypropylcellulose (HPC) andhydroxypropylmethylcellulose (HPMC), can be used. Among the acrylicpolymers, insoluble acrylate ammonio-methacrylate copolymer (Eudragit™RL100 or RS100 or Eudragit™ RL30D or RS30D), polyacrylate (Eudragit™NE30D), or methacrylic copolymers (e.g., Eudragit™ L100-55 Eudragit™L30D, Eudragit™ E100 and Eudragit™ EPO) can be used, alone, incombination, or in admixture with pH-dependent polymers. Eudragit™ E100or a mixture of Eudragit™ EPO and Eudragit™ NE30D are suitably used. Inone embodiment, the binder and the coating polymer are the same polymer.

Optionally, permeabilising agents, plasticizers, soluble agents,disintegrating agents and surfactants, can be added as coatingadditives. Suitable plasticizers include, but are not limited to,triacetine, triethylacetate, triethylcitrate (Eudraflex™),ethylphthalate, or mixtures thereof. The plasticizer is used inproportions of at most about 30%, preferably 10% by weight of thecoating polymers. Suitable soluble agents include polyols having lessthan 13 carbon atoms. Surfactants may be an anionic, nonionic, cationic,zwitterionic or amphoteric surfactant. Suitable disintegrating agentsinclude, but are not limited to, croscarmellose, available as e.g.Ac-di-sol™, crospovidone available as e.g. Kollidon CL™, and mixturesthereof.

Suitably, the coated granules according to the present invention have aparticle size distribution between about 150 μm and about 500 μm, moresuitably between about 150 μm and about 425 μm, such that at least 50%,more suitably at least 70% of the granules have a particle size rangingbetween about 150 and about 425 μm, and less than 15% of the granuleshave a particle size less than about 150 μm.

In one embodiment, the coated granules according to the inventioncomprise: from about 10% to about 95%, preferably about 40 to about 75%of granules of a compound(s) of the invention and optionally one or moreoptional additional active agents, such as those disclosed herein,sucralose from about 0.05% to about 5%, from about 5 to about 90%,suitably about 10 to about 70% and even more suitably from about 25 toabout 55% of a coating polymer, such as Eudragit™ E100, the percentagesbeing expressed by weight relative to the weight of the granules of acompound(s) of the invention, from about 0 to about 10% of apermeabilising agent, such as colloidal silica, the percentages beingexpressed by weight relative to the weight of the coating polymer.

In another embodiment, the present invention provides a solid,effervescent, rapidly dissolving dosage form of one or more compounds ofthe invention for oral administration, such as disclosed in U.S. Pat.No. 6,245,353, the disclosure of which is incorporated by referenceherein in its entirety. In such an embodiment, the effervescentformulation comprise (or consist essentially of) (a) one or morecompounds of the invention, and optionally one or more additional activeagents such as those disclosed herein, (b) an effervescent basecomprising at least one of (i) at least one of (1) an organic edibleacid and (2) a salt thereof, (ii) at least one of an alkali metal and analkaline earth metal carbonate and bicarbonate, and (c) optionally apharmaceutically acceptable auxiliary ingredient. In certain suitableembodiments, the effervescent formulations further comprise one or moretaste-masking agents, such as sucralose, and/or other taste-maskingagents described herein. The amounts of one or more compounds of theinvention, other optional active agents (e.g., anticholesterolemics,anticoagulants, anti-obesity or anti-diabetic drugs, and combinationsthereof), and sweetening agents (e.g., sucralose) in the effervescentformulations of the present invention are readily determinable by thoseof ordinary skill in the art, and include those amounts and combinationsdescribed herein. For example, the effervescent formulations of thepresent invention comprise about 0.1% to about 0.15% of one or morecompounds of the invention, optionally about 0.01% to about 10% otheractive agent(s) (or more if required), and about 0.05% to about 0.15%sucralose.

A solution or suspension of one or more compounds of the invention isformed by adding water to the soluble or dispersible effervescenttablets or soluble granules, with evolution of CO₂ gas. The resultingeffervescent solution or suspension can be ingested very easily, even bypatients who have difficulties swallowing. The rapidly disintegratingtablet can also be administered so that it directly disintegrates in themouth. A rapid release of the active ingredient is of particularimportance here, to ensure a rapid onset of action.

Effervescent agents capable of releasing CO₂, which can be used in thepresent invention, include alkali metal carbonates or alkali metalbicarbonates, such as sodium carbonate or sodium bicarbonate. Agents forinducing CO₂ release which are suitably employed are edible organicacids, or their acidic salts, which are present in solid form and whichcan be formulated with the one or more compounds of the invention activeingredient(s) and the other auxiliary ingredients (as well as any otheractive agents) to provide granules or tablets, without prematureevolution of CO₂. Edible organic acids which can be so used include forexample, tartaric acid, malic acid, fumaric acid, adipic acid, succinicacid, ascorbic acid, maleic acid or citric acid. Pharmaceuticallyacceptable acidic salts include, for example, salts of polybasic acidswhich are present in solid form and in which at least one acid functionis still present, such as sodium dihydrogen or disodium hydrogenphosphate or monosodium or disodium citrate.

In one embodiment, the present invention provides effervescentformulations of one or more compounds of the invention including theformulations and compositions described herein, having an effervescentbase comprising (a) a mixture of calcium carbonate with an organicedible acid; (b) a mixture of calcium carbonate, sodium carbonate,sodium bicarbonate and an organic edible acid; or (c) a mixture ofsodium bicarbonates, sodium carbonate and an organic edible acid.

The soluble or dispersible effervescent tablets of one or more compoundsof the invention or the soluble granules suitably comprise (orconsisting essentially of) from about 0.5 mg to about 50 mg of one ormore compounds of the invention and from about 50 mg to about 5000 mg,suitably from about 500 mg to about 3000 mg of an effervescent base,optionally, along with other active agents (such as those describedherein) and excipients, including taste-masking agents such assucralose, suitably at about 0.05% to about 5%.

The effervescent base suitably comprises from about 100 mg to about 500mg calcium ions as, for example, calcium carbonate, and from about 20 mgto about 1500 mg citric acid and/or its salts. In another embodiment,the effervescent base comprises from about 50 mg to about 2000 mg sodiumbicarbonate, from about 20 mg to about 200 mg of sodium carbonate andfrom about 20 mg to about 1500 mg citric acid and/or from about 20 mg toabout 500 mg tartaric acid.

An additional suitable composition of the effervescent base comprisesfrom about 50 mg to about 500 mg sodium bicarbonate, from about 20 mg toabout 100 mg sodium carbonate, and from about 50 mg to about 750 mgcalcium carbonate and from about 100 mg to about 1500 mg of citric acid.

The soluble/dispersible tablets can be prepared by known processes forpreparing effervescent bases, such as those disclosed in U.S. Pat. No.6,245,353, the disclosure of which is incorporated herein by referencein its entirety.

Another embodiment of the present invention is directed to aphysiologically acceptable film that is particularly well-adapted todissolve in the oral cavity of a warm-blooded animal including humans,and adhere to the mucosa of the oral cavity, to allow delivery of one ormore compounds of the invention, and optionally one or more additionalactive agents such as those described herein. Such physiologicallyacceptable films suitable for use in accordance with this aspect of thepresent invention are disclosed in U.S. Patent Application No.2004/0247648, the disclosure of which is incorporated herein byreference in its entirety.

In one such embodiment of the present invention, an orallydissolving/consumable film comprises a modified starch, one or morecompounds of the invention, and optionally, one or more additionalactive agents such as those described herein, suitably, one or moretaste-masking agents, such as sucralose, and optionally, at least onewater soluble polymer. The amounts of one or more compounds of theinvention, other optional active agents (e.g., steroids, decongestants,leukotriene antagonists, and combinations thereof), and sweeteningagents (e.g., sucralose) in the orally dissolving/consumable filmformulations of the present invention are readily determinable by thoseof ordinary skill in the art, and include those amounts and combinationsdescribed herein. For example, the orally dissolving/consumable filmformulations of the present invention comprise about 0.5 mg to about 10mg of one or more compounds of the invention, optionally about 0.50 mgto about 50 mg other active agent(s), and about 0.05% to about 0.15%sucralose.

The consumable films of the present invention may comprise one or moreof the following ingredients: water, antimicrobial agents, additionalfilm forming agents or water soluble polymers, plasticizing agents,flavorings, sulfur precipitating agents, saliva stimulating agents,cooling agents, surfactants, stabilizing agents, emulsifying agents,thickening agents, binding agents, coloring agents, triglycerides,poly(ethylene) oxides, propylene glycols, additional taste-maskingagents or sweeteners, fragrances, preservatives and the like, asdescribed in U.S. Pat. No. 6,596,298, the disclosure of which isincorporated by reference herein in its entirety.

In one such embodiment, the consumable films of the present inventioninclude a modified starch. The modified starches used in accordance withthe present invention can be prepared by mechanically, chemically orthermally modifying unmodified starches. For example, modified starchesmay be prepared by chemically treating starches to produce, for example,acid treatment starches, enzyme treatment starches, oxidized starches,cross-bonding starches, and other starch derivatives. Starches suitablefor modification to produce modified starches may be obtained fromnatural products such as corn, potatoes, tapioca as well as geneticallymodified forms of the same such as high amylose and waxy corn as well assorghum varieties.

Examples of modified starches for use in the practice of the presentinvention include, but are not limited to, modified corn starches,modified tapioca starches, acid and enzyme hydrolyzed corn and/or potatostarches, hypochlorite-oxidized starches, acid-thinned starches,ethylated starches, cross-bonded starches, hydroxypropylated tapiocastarches, hydroxypropylated corn starches, pregelatinized modifiedstarches, and the like. Preferred modified starches are selected frompregelatinized modified corn starches and pregelatinized modifiedtapioca starches.

Representative examples of commercially available modified starchesuseful in the present invention include PURE-COTE™ modified starchessuch as PURE-COTE™ B793 (a pregelatinized modified corn starch) andPURE-COTE™ B795 (a pregelatinized modified corn starch), for example,available from Grain Processing Corporation, 1600 Oregon Street,Muscatine, Iowa 52761-1494 USA.

In one such embodiment of the present invention, the modified starch ispresent in amounts ranging from about 1% to about 90% by weight, inanother embodiment about 10% to about 90% by weight, and in yet anotherembodiment from about 35% to about 80% by weight of the film.

Modified starch may be included in the film alone or optionally incombination with an additional water soluble film forming polymers suchas those selected from, for example, pullulan, hydroxypropylmethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, carboxymethyl cellulose, polyvinyl alcohol, sodiumalginate, poly(ethylene glycol), tragacanth gum, guar gum, acacia gum,arabic gum, polyacrylic acid, methylmethacrylate copolymers,carboxyvinyl polymers, amylose, high amylose starch, hydroxypropylatedhigh amylose starch, pectin, dextrin, chitin, chitosan, levan, elsinan,collagen, gelatin, zein, gluten, soy protein isolate, whey proteinisolate, casein and combinations thereof. A preferred water solublepolymer is pullulan. The amount of the water soluble polymer typicallyis up to about 99% by weight, suitably up to about 80% by weight, moresuitably up to about 50% by weight, and most suitably up to about 40% byweight of the film.

Suitable formulations for oral and/or parenteral administration includeaqueous solutions of one or more of the compounds of the invention, andoptionally one or more additional active pharmaceutical ingredients, inwater-soluble form, for example, water-soluble salts and alkalinesolutions. In addition, suspensions of the active ingredient(s) asappropriate oily injection suspensions may be administered. Suitablelipophilic solvents or vehicles include fatty oils, for example, sesameoil, or synthetic fatty acid esters, for example, ethyl oleate ortriglycerides or poly(ethylene glycol)-400. Aqueous injectionsuspensions may optionally also comprise substances which increase theviscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol, and/or dextran. Optionally, the suspension may alsocontain one or more stabilizers, one or more preservatives (e.g., sodiumedetate, benzalkonium chloride, and the like), and/or other componentscommonly used in formulating pharmaceutical compositions.

Suitable topical pharmaceutical compositions of the invention areformulated preferably as oils, creams, lotions, ointments and the likeby choice of appropriate carriers. Such compositions of the inventiontherefore comprise one or more compounds of the invention, optionallyone or more additional active pharmaceutical ingredients, and one ormore carriers suitable for use in preparing such pharmaceuticalcompositions for topical administration. Suitable such carriers includevegetable or mineral oils, white petrolatum (white soft paraffin),branched chain fats or oils, animal fats and high molecular weightalcohol (greater than C12). The preferred carriers are those in whichthe active pharmaceutical ingredient(s) are soluble. Emulsifiers,stabilizers, humectants and antioxidants may also be included, as wellas agents imparting color or fragrance, if desired. Additionally, one ormore transdermal penetration enhancers can be employed in these topicalformulations. Non-limiting examples of suitable such enhancers can befound in U.S. Pat. Nos. 3,989,816 and 4,444,762, which are incorporatedbe reference herein in their relevant parts.

Creams are preferably formulated from a mixture of mineral oil,self-emulsifying beeswax and water in which mixture the activeingredient, dissolved in a small amount of an oil such as almond oil, isadmixed. A typical example of such a cream is one which includes about40 parts water, about 20 parts beeswax, about 40 parts mineral oil andabout 1 part almond oil.

Ointments may be formulated by mixing a solution of the activeingredient in a vegetable oil such as almond oil with warm soft paraffinand allowing the mixture to cool. A typical example of such an ointmentis one which includes about 30% almond oil and about 70% white softparaffin by weight.

Lotions may be conveniently prepared by dissolving the activeingredient, in a suitable high molecular weight alcohol such aspropylene glycol or poly(ethylene glycol).

Suitable liquid pharmaceutical compositions for ocular administrationcomprise (or consisting essentially of) a therapeutically effective doseof one or more compounds of the invention, and one or morepharmaceutically acceptable carriers or excipients, wherein at least oneof the pharmaceutically acceptable carriers or excipients is sucralose,wherein the composition is free, or substantially free of preservatives,and wherein the composition is provided in a single unit-dose container.Suitably, the amount of one or more compounds of the invention in suchliquid, unit-dose pharmaceutical compositions is about 0.05% to about0.15% by weight and the amount of sucralose in such liquid, unit-dosepharmaceutical compositions is about 0.05% to about 0.15% by weight.Suitable unit-dose containers include, but are not limited to, highdensity polyethylene containers, for example, high density polyethylenecontainers produced using a blow-fill-seal manufacturing technique witha volume capacity of about 1 mL.

Suitable liquid pharmaceutical compositions for nasal administration inunit-dose or multi-dose configurations, comprising (or consistingessentially of) a therapeutically effective dose of one or morecompounds of the invention, and one or more pharmaceutically acceptablecarriers or excipients, wherein at least one of the pharmaceuticallyacceptable carriers or excipients is sucralose, wherein the compositionis free, or substantially free of preservatives, and wherein thecomposition is provided in either a unit-dose or multi-dose container.Suitably, the amount of one or more compounds of the invention in suchliquid, unit-dose or multi-dose pharmaceutical compositions is about0.05% to about 0.15% by weight and the amount of sucralose in suchliquid, unit-dose or multi-dose pharmaceutical compositions is about0.05% to about 0.15% by weight. Suitable unit-dose or multi-dosecontainers include, but are not limited to, high density polyethylenebottles with a volume capacity of about 1 ml to 10 mL fitted with aspray pump specifically designed for use with preservative freeformulations.

The present invention also provides inhalable powder pharmaceuticalcompositions comprising (or consisting essentially of), atherapeutically effective dose of one or more compounds of theinvention, and one or more pharmaceutically acceptable carriers orexcipients, wherein the compound(s) of the invention are in the form ofmicronized particles and wherein at least one of the pharmaceuticallyacceptable carriers or excipients is sucralose, for example, micronizedparticles of sucralose. Suitable such inhalable powder pharmaceuticalcompositions comprise micronized particles of one or more compounds ofthe invention with an average particle size of about 1 μm to about 5 μm,and micronized particles of sucralose with an average particle size ofabout 1 μm to about 20 μm. Such inhalable powder pharmaceuticalcompositions of the present invention can be formulated for pulmonarydelivery using, for example, a dry powder inhaler. Suitably, the amountof one or more compounds of the invention in such inhalable powderpharmaceutical compositions is about 0.1% to about 20.0% by weight andthe amount of sucralose in such inhalable powder pharmaceuticalcompositions is about 0.05% to about 20.0% by weight.

The present invention also provides inhalable spray pharmaceuticalcompositions comprising (or consisting essentially of), a suitableconcentration to provide a therapeutically effective dose of one or morecompounds of the invention, and one or more pharmaceutically acceptablecarrier, stabilizer or excipient, wherein the compound(s) of theinvention is(are) in a solution form and wherein at least one of thepharmaceutically acceptable carriers or excipients is sucralosedissolved in the solution. Such inhalable spray pharmaceuticalcompositions when used with a suitable device provide a fine spray ofthe components (including active and non-active components) having anaverage particle size of about 1 μm to about 5 μm. Such inhalable spraypharmaceutical compositions of the present invention can be formulatedfor pulmonary delivery using, for example, a suitable device or inhaler.Suitably the amount of one or more compounds of the invention in suchinhalable spray pharmaceutical compositions is about 0.1% to about 10%by weight and the amount of sucralose in such inhalable spraypharmaceutical compositions is about 0.05% to about 0.15% by weight.

Liquid dosage forms for nasal, ocular or oral administration includepharmaceutically acceptable emulsions, solutions, suspensions, syrupsand elixirs. In addition to the active one or more compounds of theinvention, the liquid dosage forms may contain inert diluents and/orsolvents commonly used in the art. Water is the solvent of choice forthe formulations of the invention; however, combinations of water withother physiologically acceptable solvents as required are alsosatisfactory for use. Other solvents, solubilizing agents andemulsifiers suitable for use in place of, or in addition to, waterinclude but are not limited to saturated aliphatic mono- and polyvalentalcohols which contain 2-6 carbon atoms (including, but not limited to,ethanol, 1,2-propylene glycol, sorbitol, and glycerine), polyglycolssuch as poly(ethylene glycols), and surfactants/emulsifiers like thefatty acid esters of sorbitan, and mixtures thereof. Oils, inparticular, cottonseed, peanut, or corn oils, may also be added to thecompositions. The combination of the additional solvents in the aqueoussolution should preferably not exceed about 15% (w/v) of the totalcomposition. Besides inert diluents, the oral compositions can alsoinclude adjuvants such as wetting agents, emulsifying and suspendingagents (e.g., microcrystalline cellulose, sodium carboxymethylcellulose, hypromellose, carbopol and the like), surfactants,sweetening, flavoring, and perfuming agents, including those describedin further detail herein below. Liquid dosage forms that provide theactive ingredient in suspension may comprise, in addition to the activeone or more compounds of the invention, one or more suspending agentssuch as microcrystalline cellulose, magnesium aluminum silicate,bentonite, agar-agar, hypromellose, sodium carboxymethyl cellulose,carbopol/carbomer, pectin, acacia, tragacanth or their mixtures.

Certain liquid compositions of the invention may further comprise one ormore preservatives and/or one or more stabilizers. Preservatives thatare suitable for use in the compositions of the invention include, butare not limited to, edetic acid and their alkali salts such as disodiumEDTA (also referred to as “disodium edetate” or “the disodium salt ofedetic acid”) and calcium EDTA (also referred to as “calcium edetate”),benzyl alcohol, methylparaben, propylparaben, butylparaben,chlorobutanol, phenylethyl alcohol, benzalkonium chloride, thimerosal,propylene glycol, sorbic acid, and benzoic acid derivatives. Thepreservatives should be used at a concentration of from about 0.001% toabout 0.5% (w/v) in the final composition. The combination ofbenzalkonium chloride, used at a concentration of from about 0.001% toabout 0.5% or preferably from about 0.005% to about 0.1% (w/v), andedetic acid (as a disodium salt), used at a concentration of from about0.005% to about 0.1% (w/v), are suitable preservative/stabilizercombination used in the compositions of the present invention.

Certain compositions of the invention may further comprise one or moresolubility-enhancing agents that are used to improve the solubility ofthe one or more compounds of the invention used as an active ingredient.Solubility-enhancing agents that are suitable for use in thecompositions of the invention include, but are not limited to,polyvinylpyrrolidone (preferably grades 25, 30, 60, or 90), poloxamer,polysorbate 80, sorbitan monooleate 80, and poly(ethylene glycols)(molecular weights of 200 to 600).

Certain compositions of the invention may further comprise one or moreagents that are used to render the composition isotonic, particularly inthose compositions in which water is used as a solvent. Such agents areparticularly useful in compositions formulated for nasal or ocularapplication, since they adjust the osmotic pressure of the formulationsto the same osmotic pressure as nasal or ocular secretions. Agents thatare suitable for such a use in the compositions of the inventioninclude, but are not limited to, sodium chloride, sorbitol, propyleneglycol, dextrose, sucrose, and glycerine, and other isotonicity agentsthat are known in the art (see, e.g., Reich et al., “Chapter 18:Tonicity, Osmoticity, Osmolality and Osmolarity,” in: Remington: TheScience and Practice of Pharmacy, 20^(th) Edition, Lippincott Williamsand Wilkins, Philadelphia, Pa. (2000)).

It is desirable that the compositions of the present invention that areto be administered in liquid form (including intranasally, orally orocularly applied formulations) have a pH of about 4.5 to about 7.4, andpreferably have a pH of about 5.5 to 7.1, for physiological reasons.Accordingly, in additional embodiments, the compositions of theinvention may further comprise one or more buffering agents orcombinations thereof, that are used to adjust and/or maintain thecompositions into the desired pH range. Adjustment of pH or bufferingagents that are suitable for use in the compositions of the inventioninclude, but are not limited to, citric acid, sodium citrate, sodiumphosphate (dibasic, heptahydrate form), and boric acid or equivalentconventional buffers, or combinations thereof. The appropriate amountsof buffers and buffering agents, or combinations thereof, that are to beused in the compositions of the invention are readily determined bythose of ordinary skill without undue experimentation, particularly inview of the guidance contained herein and in standard formularies suchas the United States Pharmacopoeia, Remington: The Science and Practiceof Pharmacy, and the like, the disclosures of which are incorporatedherein by reference in their entireties.

In certain embodiments, the liquid formulations of the invention,particularly those that are to be administered intranasally, ocularly,or orally, further comprise one or more taste-masking agents, one ormore flavoring agents, and/or one or more sweetening agents, or acombination of such agents. Non-limiting examples of such substancesinclude sucralose (about 0.001 to about 1%), sucrose (about 0.5 to about10%), saccharin (including the salt forms: sodium, calcium, etc.) (about0.01 to about 2%), fructose (about 0.5 to about 10%), dextrose (about0.5 to about 10%), corn syrup (about 0.5 to about 10%), aspartame (about0.01 to about 2%), acesulfame-K (about 0.01 to about 2%), xylitol (about0.1 to about 10%), sorbitol (about 0.1 to about 10%), erythritol (about0.1 to about 10%), ammonium glycyrrhizinate (about 0.01 to about 4%),thaumatin (Talin™) (about 0.01 to about 2%), neotame (about 0.01 toabout 2%) mannitol (about 0.5 to about 5%), menthol (about 0.01 to about0.5%), eucalyptus oil (about 0.01 to about 0.5%), camphor (about 0.01 toabout 0.5%), natural and/or artificial flavors such as ArtificialCustard Cream Flavor #36184 from International Flavors and Fragrances,Inc. (New York, N.Y.) (about 0.01 to about 1.0%), and the like.Sucralose, an intense sweetener marketed for food and beverage use asSPLENDA® by McNeil Nutritionals LLP (Fort Washington, Pa.), isespecially effective as a sweetening and taste-masking agent in thecompositions of the present invention, particularly when used atconcentrations of from about 0.001% to about 1%, preferably atconcentrations of from about 0.01% to about 0.5%, and more preferably atconcentrations of from about 0.02% to about 0.2%, and most preferablyfrom about 0.05% to about 0.15% (e.g., about 0.05%, about 0.06%, about0.07%, about 0.08%, about 0.09%, about 0.10%, about 0.11%, about 0.12%,about 0.13%, about 0.14%, or about 0.15%), of the total composition.Sucralose has been shown to be useful as a taste modifying agent in oraldelivery of certain pharmaceutical compositions, for example in sorethroat spray products (see U.S. Pat. No. 6,319,513), oral suspensions(see U.S. Pat. Nos. 5,658,919 and 5,621,005), solid dosage forms (seeU.S. Pat. No. 6,149,941), quick melt dosage forms (see U.S. Pat. No.6,165,512) and mucosal delivery (see U.S. Pat. No. 6,552,024), but hasnot heretofore been shown to be useful in intranasally or ocularlyapplied compositions such as those of the present invention. Additionalsuch compositions of the invention may comprise one or more additionaltaste-masking or flavoring agents such as those described herein, forexample menthol at a concentration of from about 0.01% to about 1%,preferably at a concentration of from about 0.05% to about 0.1%.Suitable compositions of the invention include, for example, about0.1%-0.15% of one or more compounds of the invention and about0.05%-0.15% sucralose, for example, about 0.1% one or more compounds ofthe invention and about 0.05%-0.15% sucralose, or about 0.125%-0.15% oneor more compounds of the invention and about 0.05%-0.15% sucralose, orabout 0.10% one or more compounds of the invention and about 0.15%sucralose, or about 0.15% one or more compounds of the invention andabout 0.15% sucralose.

In further embodiments, the present invention provides formulations andcompositions for pulmonary delivery of one or more compounds of theinvention, and optionally, one or more additional active agents, such asthose described herein. For example, inhalable preparations comprisingone or more compounds of the invention, and optionally, one or moreadditional active agents such as those described herein, can beproduced.

Inhalable preparations include inhalable powders, propellant-containingmetering aerosols or propellant-free inhalable solutions Inhalablepowders according to the invention containing one or more compounds ofthe invention, and optionally one or more additional active ingredientsincluding those described herein, may comprise the active ingredients ontheir own, or a mixture of the active ingredients with physiologicallyacceptable excipients. In certain such embodiments, the inhalableformulas comprise the compositions of the present invention in aninhalable form. Within the scope of the present invention, the termpropellant-free inhalable solutions also includes concentrates orsterile inhalable solutions ready for use. The preparations according tothe invention may comprise one or more compounds of the invention andoptionally one or more additional active ingredients including thosedescribed herein, in one formulation, or in two or more separateformulations.

Physiologically acceptable excipients that may be used to prepare theinhalable powders according to the present invention include, but arenot limited to, monosaccharides (e.g., glucose or arabinose),disaccharides (e.g., lactose, saccharose, maltose), oligo- andpolysaccharides (e.g., dextran), polyalcohols (e.g., sorbitol, mannitol,xylitol), salts (e.g., sodium chloride, calcium carbonate) or mixturesof these excipients with one another. Suitably, mono- or disaccharidesare used, for example, lactose or glucose in the form of their hydrates.Lactose and lactose monohydrate represent exemplary excipients.Excipients for use in the inhalable preparations can have an averageparticle size of up to about 250 μm, suitably between about 10 μm andabout 150 μm, most suitably between about 15 μm and about 80 μm. Incertain such embodiments, finer excipient fractions can be added with anaverage particle size of about 1 μm to about 9 μm. These finerexcipients are also selected from the group of possible excipientsdisclosed throughout. Finally, in order to prepare the inhalable powdersaccording to the present invention, micronised active ingredients (e.g.,one or more compounds of the invention and optionally one or moreadditional agents described throughout), suitably with an averageparticle size of about 0.5 μm to about 10 μm, more suitably from about 1μm to about 5 μm, are added to the excipient mixture. Processes forproducing the inhalable powders according to the present invention bygrinding and micronizing and by finally mixing the ingredients togetherare routine and well known to those of ordinary skill in the art. Theinhalable powders according to the present invention can be prepared andadministered either in the form of a single powder mixture whichcontains one or more compounds of the invention and optionally one ormore additional active agents such as those described herein, or in theform of separate inhalable powders, in which one powder contains onlyone or more compounds of the invention, and another powder contains oneor more additional active agents such as those described herein. Methodsfor preparing the inhalable powders of the present invention, as well asdevices for their delivery, are disclosed in U.S. Pat. Nos. 6,696,042and 6,620,438; U.S. Published Patent Application Nos. 2002/0009418,2005/0121032, 2005/0121027 and 2005/0123486, the disclosures of each ofwhich are incorporated herein by reference in their entireties.

The inhalable powders according to the present invention may beadministered using inhalers well known in the art Inhalable powdersaccording to the present invention which contain a physiologicallyacceptable excipient in addition to the active agents may beadministered, for example, by means of inhalers which deliver a singledose from a supply using a measuring chamber as described in U.S. Pat.No. 4,570,630, or by other means as described in U.S. Pat. Nos.5,035,237 and 4,811,731, the disclosures of which are incorporated byreference herein in their entireties. The inhalable powders of thepresent invention can also be administered by dry powder inhalers (DPIs)or pre-metered DPIs (see e.g., U.S. Pat. Nos. 6,779,520, 6,715,486 and6,328,034, the disclosures of each of which are incorporated herein byreference in their entireties). Suitably, the inhalable powdersaccording to the present invention which contain physiologicallyacceptable excipients in addition to the active agents are packed intocapsules (to produce so-called inhalettes) which are used in inhalers asdescribed, for example, in U.S. Pat. No. 5,947,118, the disclosure ofwhich is incorporated herein by reference in its entirety. An additionalDPI that can be used with the powder formulations of the presentinvention is the Novalizer® by Sofotec (Bad Homburg, Germany). Adescription of this DPI, as well as methods to formulate powders for usein it, are disclosed in U.S. Pat. Nos. 5,840,279; 5,881,719; 6,071,498;and 6,681,768, the disclosures of which are incorporated herein byreference in their entireties.

According to another embodiment of the present invention, inhalationaerosols containing propellant gas comprising (or consisting essentiallyof) one or more compounds of the invention, and optionally, one or moreadditional active ingredients such as those described herein, dissolvedin a propellant gas or in dispersed form, can be produced. One or morecompounds of the invention, and one or more optional active ingredients,such as those disclosed herein, may be present in separate formulationsor in a single preparation, in which all active ingredients are eachdissolved, each dispersed, or one or more active components aredissolved and any others are dispersed. The propellant gases which maybe used to prepare the inhalation aerosols according to the inventionare known in the art. Suitable propellant gases include, but are notlimited to, hydrocarbons such as n-propane, n-butane or isobutane andhalohydrocarbons such as fluorinated derivatives of methane, ethane,propane, butane, cyclopropane or cyclobutane. The propellant gases maybe used on their own or in mixtures thereof. Particularly suitablepropellant gases are halogenated alkane derivatives selected from TG134aand TG227.

The propellant-driven inhalation aerosols according to the presentinvention may also contain other ingredients such as co-solvents,stabilizers, surfactants, antioxidants, lubricants and pH adjusters. Allof these ingredients, and suitable commercial sources thereof, are wellknown in the art.

The inhalation aerosols containing propellant gas according to thepresent invention may contain up to about 5 wt. % of active substances(or more if required). Aerosols according to the invention contain, forexample, about 0.002 wt. % to about 5 wt. %, about 0.01 wt. % to about 3wt. %, about 0.015 wt. % to about 2 wt. %, about 0.1 wt. % to about 2wt. %, about 0.5 wt. % to about 2 wt. %, or about 0.5 wt. % to about 1wt. % of active substances (e.g., one or more compounds of the inventionand optionally one or more additional active agents such as thosedescribed herein).

In embodiments where the active substance(s) are present in dispersedform, the particles of active substance(s) suitably have an averageparticle size of up to about 10 μm, suitably from about 0.1 μm to about5 μm, more suitably from about 1 μm to about 5 μm.

Propellant-driven inhalation aerosols according to certain suchembodiments of the present invention may be administered using inhalersknown in the art (e.g., MDIs: metered dose inhalers, see e.g., U.S. Pat.Nos. 6,380,046, 6,615,826 and 6,260,549, the disclosures of each ofwhich are incorporated herein by reference in their entireties).Accordingly, in another aspect, the present invention providespharmaceutical compositions in the form of propellant-driven aerosolscombined with one or more inhalers suitable for administering theseaerosols. In addition, the present invention provides inhalers which arecharacterized in that they contain the propellant gas-containingaerosols described throughout. The present invention also providescartridges which are fitted with a suitable valve and can be used in asuitable inhaler and which contain one or more of the propellantgas-containing inhalation aerosols described throughout. Suitablecartridges and methods of filling these cartridges with the inhalableaerosols containing propellant gas according to the invention are knownin the art.

In another embodiment, the present invention provides propellant-freeinhalable formulations, such as solutions and suspensions, comprising(or consisting essentially of) one or more compounds of the inventionand optionally one or more additional active agents such as thosedescribed herein. Suitable solvents for use in such embodiments includeaqueous and alcoholic solvents, suitably an ethanolic solution. Thesolvents may be water on its own or a mixture of water and ethanol. Therelative proportion of ethanol compared with water suitably is up toabout 70 percent by volume, more suitably up to about 60 percent byvolume, or up to about 30 percent by volume. The remainder of the volumeis made up of water. The solutions or suspensions containing one or morecompounds of the invention and optionally one or more additional activeagents, such as those described herein, separately or together, areadjusted to a pH of 2 to 7, using suitable acids or bases. The pH may beadjusted using acids selected from inorganic or organic acids. Examplesof suitable inorganic acids include hydrochloric acid, hydrobromic acid,nitric acid, sulfuric acid and phosphoric acid. Examples of suitableorganic acids include ascorbic acid, citric acid, malic acid, tartaricacid, maleic acid, succinic acid, fumaric acid, acetic acid, formicacid, propionic acid, etc. Exemplary inorganic acids includehydrochloric and sulfuric acids. It is also possible to use the acidswhich have already formed an acid addition salt with one or more of theactive substances. Exemplary organic acids include ascorbic acid,fumaric acid and citric acid. If desired, mixtures of the above acidsmay be used, particularly in the case of acids which have otherproperties in addition to their acidifying qualities, e.g., asflavorings, antioxidants or complexing agents, such as citric acid orascorbic acid, for example. Hydrochloric acid can be used to adjust thepH.

Co-solvents and/or other excipients may be added to the propellant-freeinhalable formulations of the present invention. Suitable co-solventsare those which contain hydroxyl groups or other polar groups, e.g.,alcohols—such as isopropyl alcohol, glycols—such as propylene glycol,poly(ethylene glycol), polypropylene glycol), glycol ether, glycerol,poly(oxyethylene alcohols) and poly(oxyethylene fatty acid esters). Theterms excipients and additives in this context denote anypharmacologically acceptable substance which is not an active substancebut which can be formulated with the active substance or substances inthe pharmacologically suitable solvent in order to improve thequalitative properties of the active substance formulation. Suitably,these substances have no pharmacological effect or, in connection withthe desired therapy, no appreciable or at least no undesirablepharmacological effect. The excipients and additives include, forexample, surfactants such as soy lecithin, oleic acid, sorbitan esters,such as polysorbates, polyvinylpyrrolidone, other stabilizers,complexing agents, antioxidants and/or preservatives which prolong theshelf life of the finished pharmaceutical formulation, flavorings,vitamins and/or other additives known in the art. The additives alsoinclude pharmacologically acceptable salts such as sodium chloride asisotonic agents.

Exemplary excipients include antioxidants such as ascorbic acid, vitaminA, vitamin E, tocopherols and similar vitamins and provitamins occurringin the human body.

Preservatives may be used to protect the inhalable formulationsdisclosed herein from contamination with pathogens. Suitablepreservatives are those which are known in the art, particularly cetylpyridinium chloride, benzalkonium chloride or benzoic acid or benzoatessuch as sodium benzoate in the concentration known from the prior art.The preservatives mentioned above are suitably present in concentrationsof up to about 50 mg/100 ml, more suitably between about 5 and about 20mg/100 ml. Alternatively, the inhalable formulations can be preparedwithout preservatives, for example, in unit-dose forms, such asdescribed herein.

The propellant-free inhalable formulations according to the presentinvention can be administered using inhalers of the kind which arecapable of nebulizing a small amount of a liquid formulation in thetherapeutic dose within a few seconds to produce an aerosol suitable fortherapeutic inhalation. Suitable inhalers are those in which a quantityof less than about 100 μL, less than about 50 μL, or between about 10 μLand about 30 μL of active substance solution can be nebulized in onespray action to form an aerosol with an average particle size of lessthan about 20 μm, suitably less than about 10 μm, in such a way that theinhalable part of the aerosol corresponds to the therapeuticallyeffective quantity.

Suitable apparatuses for propellant-free delivery of a metered quantityof a liquid pharmaceutical composition according to the presentinvention are described for example in U.S. Pat. Nos. 5,497,944;5,662,271; 5,964,416; 6,402,055; 6,497,373; 6,726,124; and 6,918,547,the disclosures of which are incorporated herein by reference in theirentireties. In another embodiment, the present invention providespharmaceutical formulations in the form of propellant-free inhalableformulations, such as solutions or suspensions, as described herein,combined with a device suitable for administering such formulations.

The propellant-free inhalable formulations, e.g., solutions orsuspensions, according to the present invention may take the form ofconcentrates or sterile inhalable solutions or suspensions ready foruse. Formulations ready for use may be produced from the concentrates,for example, by the addition of isotonic saline solutions. Sterileformulations ready for use may be administered using energy-operatedfixed or portable nebulizers which produce inhalable aerosols by meansof ultrasound or compressed air by the Venturi principle or otherprinciples.

The present invention also provides fine particle dosages of one or morecompounds of the invention and optionally one or more additional activeagents such as those described herein. A delivered fine particle dose(FPD) of one or more compounds of the invention administered byinhalation herein is not limited, and may generally be in a range fromabout 1 to about 50 μg, including about 5, 10, 15, 20, 30 and 40 μg. Thecorrect metered dose loaded into an inhaler to be used for the purposeof administration can be adjusted for predicted losses such as retentionand more or less efficient de-aggregation of the inhaled dose.

Excipient particles having a physical median particle size larger thanabout 25 □m and having a very narrow particle size distribution withgenerally less than 5% of the particles by mass being below 10 μmgenerally show good flow properties, and are suitable for use inmixtures together with one or more compounds of the invention andoptionally one or more additional active agents, such as those describedherein. For inhalation purposes, carrier particles having a mass medianparticle size in a range from about 10 to about 250 μm are typicallyselected, including about 30, 50, 70, 100, 130, 160, 190, and 220 μm.The median particle size chosen within this range depends on manyfactors, e.g. type of carrier substance, degree of powder flowability tobe attained, type of inhaler and ease of de-aggregation duringinhalation of the resulting medicament. Commercial grades of Respitosare available (lactose monohydrate from DMV of several defined particlesize distributions up to 400 μm) suitable as particular excipients to beused in formulations containing one or more compounds of the invention,e.g. grade SV003. Uniform homogeneous one or more compounds of theinvention powder formulations having a physical median particle sizedown to about 10 □m can also provide good flow properties when theparticles have been modified to have a very smooth surface, therebyimproving the flow properties of the formulation.

A practical lower limit for volumetric dose forming for such inhalablepowder formulations is in a range of about 0.5 to 1 mg. Smaller dosescan be difficult to produce and still maintain a low relative standarddeviation between doses in the order of 10%. Typically, though, dosemasses range from about 1 to 10 mg.

Suitable excipients for inclusion in the one or more compounds of theinvention powder formulations include, but are not limited to,monosaccarides, disaccarides, polylactides, oligo- and polysaccarides,polyalcohols, polymers, salts or mixtures from these groups, e.g.glucose, arabinose, lactose, lactose monohydrate, lactose anhydrous(i.e., no crystalline water present in lactose molecule), saccharose,maltose, dextran, sorbitol, mannitol, xylitol, sodium chloride andcalcium carbonate.

Excipients for use with one or more compounds of the invention andoptionally one or more additional active agents, such as those describedherein, generally are selected from among excipients which have goodmoisture qualities in the sense that the substance will not adverselyaffect the active agent fine particle dose (FPD) for the shelf life ofthe product regardless of normal changes in ambient conditions duringstorage. Suitable “dry” excipients are well known in the art and includethose disclosed herein. For example, lactose can be selected as a dryexcipient, or lactose monohydrate can be used in a formulation with oneor more compounds of the invention (and optionally one or moreadditional active agents, such as those described herein). Lactose hasthe inherent property of having a low and constant water sorptionisotherm. Excipients having a similar or lower sorption isotherm canalso be used.

As discussed throughout, and in a further aspect of the presentinvention, one or more compounds of the invention may be mixed orformulated with one or more additional active agents such as thosedescribed herein in the dry powder or other inhalable formulations. Thepresent invention also encompasses the use of one or more compounds ofthe invention where a combination of one or more compounds of theinvention with other agents, such as those described herein, constitutea formulation from which metered doses are then produced, filled andsealed into dry, moisture-tight, high barrier seal containers intendedfor insertion into a DPI to be administered according to a particulardosing regime or as needed by the user. Suitable additional activeagents include those disclosed throughout, for example,anticholesterolemics, anticoagulants, anti-obesity or anti-diabeticdrugs.

A sealed, dry, high barrier container can be loaded with a powder formof one or more compounds of the invention and optionally one or moreadditional active agents, such as those described herein, in the form ofa blister and may comprise a flat dose bed or a formed cavity inaluminum foil or a molded cavity in a polymer material, using a highbarrier seal foil against ingress of moisture, e.g. of aluminum or acombination of aluminum and polymer materials. The sealed, dry, highbarrier container may form a part of an inhaler device or it may form apart of a separate item intended for insertion into an inhaler devicefor administration of pre-metered doses.

The present invention also provides inhalable spray pharmaceuticalcompositions comprising (or consisting essentially of), a suitableconcentration to provide a therapeutically effective dose of one or morecompounds of the invention, and one or more pharmaceutically acceptablecarrier, stabilizer or excipient, wherein the one or more compounds ofthe invention is in a solution form and wherein at least one of thepharmaceutically acceptable carriers or excipients is sucralosedissolved in the solution. Such inhalable spray pharmaceuticalcompositions when used with a suitable device provide a fine spray ofthe components (including active and non-active components) having anaverage particle size of about 1 μm to about 5 μm. Such inhalable spraypharmaceutical compositions of the present invention can be formulatedfor pulmonary delivery using, for example, a suitable device or inhaler.Suitably the amount of one or more compounds of the invention in suchinhalable spray pharmaceutical compositions is about 0.1% to about 10%by weight and the amount of sucralose in such inhalable spraypharmaceutical compositions is about 0.05% to about 0.15% by weight,though other suitable amounts will readily be determined by theordinarily skilled artisan.

In certain embodiments, a pharmaceutical composition comprising acompound of the invention and one or more additional therapeutic agentsare administered to a patient.

In certain embodiments, compounds of the invention and one or moreadditional therapeutic agents are administered to a patient in separatecompositions under one or more of the following conditions: at differentperiodicities, at different durations, at different concentrations, bydifferent administration routes, etc. In one such embodiment, thecompound of Formula I is administered prior to the one or moreadditional therapeutic agents, e.g., 0.5, 1, 2, 3, 4, 5, 10, 12, or 18hours, 1, 2, 3, 4, 5, or 6 days, 1, 2, 3, or 4 weeks prior to theadministration of the therapeutic agent(s). In another such embodiment,the compound is administered after the one or more additionaltherapeutic agents, e.g., 0.5, 1, 2, 3, 4, 5, 10, 12, or 18 hours, 1, 2,3, 4, 5, or 6 days, 1, 2, 3, or 4 weeks after the administration of thetherapeutic agent(s). In another such embodiment, the compound ofFormula I and the one or more additional therapeutic agents areadministered concurrently but on different schedules, e.g., the compoundof Formula I is administered daily while the one or more additionaltherapeutic agents are administered once every two days, once everythree days, once every four days, once every five days, once every sixdays, once a week, once every two weeks, once every three weeks, onceevery four weeks, etc.

General Synthesis of β-Hydroxy-γ-Aminophosphonates

Scheme 1 depicts a general synthesis of β-hydroxy-γ-aminophosphonate andβ-hydroxy-γ-aminophosphonate analogs of Formula A, wherein R is selectedfrom the group consisting of hydrogen and lower alkyl, R¹ is selectedfrom the group consisting of hydrogen, optionally substituted alkyl,aralkyl, optionally substituted cycloalkyl, optionally substitutedalkenyl, optionally substituted alkynyl, optionally substituted aryl,optionally substituted heteroaryl and optionally substitutedheterocyclo, R^(2a) and R^(3a) are independently selected from the groupconsisting of optionally substituted alkyl, aralkyl and optionallysubstituted aryl, R⁴ is selected from the group consisting of hydrogen,optionally substituted alkyl and COR⁵, R⁵ is selected from the groupconsisting of optionally substituted alkyl, aralkyl, optionallysubstituted cycloalkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted aryl, optionally substitutedheteroaryl and optionally substituted heterocyclo, and X⁻ is apharmaceutically acceptable anion.

In reaction (i.e., rxn) 1, the amine functionality of a racemic aminoacid a (e.g., D,L-leucine) is protected and the carboxylic acid isconverted to an ester to give b, wherein R⁶ is an amine protecting groupand R⁷ is selected from the group consisting of hydrogen and amineprotecting group, or R⁶ and R⁷ taken together represent an amineprotecting group, and R⁸ is selected from the group consisting ofoptionally substituted alkyl, aralkyl, and optionally substituted aryl.Suitable amine protecting groups and synthetic methods used to introduceamine protecting groups are well known in the art of organic synthesis.See, for example, Greene and Wuts, “Protective Groups in OrganicSynthesis,” 3rd Ed., pp. 17-245 (J. Wiley & Sons, 1999). Likewise,synthetic methods used to esterify a carboxylic acid are well known inthe art. The amine of a may be protected before esterification, afteresterification or simultaneously with esterification. In an exemplaryembodiment, the amino acid a is treated with benzyl bromide in thepresence of potassium carbonate to give b, wherein R⁶, R⁷ and R⁸ areeach benzyl. In certain embodiments, the reaction is carried in asolvent system comprising a C₁-C₄ alcohol (e.g., methanol) and water oracetonitrile and water. In certain embodiments, the reaction is carriedout at reflux temperature. In certain embodiments, b is used in the nextsynthetic step without purification.

In reaction 2, b is condensed with Li⁺(⁻CH₂P(O)(OR^(2a))(OR^(3a)) togive a β-keto-γ-aminophosphonate c. In certain embodiments, R^(2a) andR^(3a) are each lower alkyl (e.g., methyl) or aralkyl (e.g., benzyl). Incertain embodiments, the condensation is carried out in tetrahydrofuranat temperature ranging from about −78° C. to about −20° C., e.g., about−50° C. The product c can be used in the next synthetic reaction withoutfurther purification.

In reaction 3, the oxo group, i.e., C═O, of a β-keto-γ-aminophosphonatec is reduced to give a β-hydroxy-γ-aminophosphonate d. In certainembodiments, the reducing agent is a borohydride reducing agent, e.g.,sodium borohydride. In certain embodiments, the reaction is carried outin a solvent system comprising a C₁-C₄ alcohol (e.g., methanol) andtetrahydrofuran. In a further embodiment, the solvent system comprisesabout 5% to about 15% methanol in tetrahydrofuran (v/v), e.g., about 10%methanol and about 90% tetrahydrofuran (v/v). In certain embodiments,the reduction is carried out a temperature from about −30° C. to about−20° C., e.g., about −10° C. to about 5° C. In certain embodiments, acompound of Formula d is used in the next synthetic reaction withoutpurification.

In reaction 4, the hydroxy group of β-hydroxy-γ-aminophosphonate d isalkylated or acylated to give e, wherein R⁴ is optionally substitutedalkyl or COR⁵, respectively. The optionally substituted alkyl group canbe introduced via reaction of d with R⁴L² wherein L² is a leaving group,e.g., methyl triflate, i.e., MeOSO₂CF₃. In certain embodiments, thereaction is carried out in an inert organic solvent, e.g.,tetrahydrofuran, dichloromethane, acetonitrile, dimethylformamide, etc.The COR⁵ group can be introduced via reaction of d with R⁵COL¹ whereinL¹ is a leaving group, e.g., acetyl chloride, i.e., MeCOCl.Alternatively, β-hydroxy-γ-aminophosphonate d can be used in the nextsynthetic step without further chemical modification, i.e., R⁴ ishydrogen.

In reaction 5, the amine protecting group(s) of e is(are) removed, i.e.,R⁶ is an amine protecting group and R⁷ is hydrogen, or R⁶ and R⁷ areboth an amine protecting groups, or R⁶ and R⁷ taken together representan amine protecting group, to give amine f. In certain embodiments, R⁶and R⁷ are benzyl. In a further embodiment, the benzyl groups areremoved, i.e., the amine is deprotected, under an atmosphere of hydrogengas using palladium on carbon as the catalyst. In a further embodimentthe deprotection is carried out in a solvent selected from the groupconsisting of C₁-C₄ alcohol (e.g., methanol) tetrahydrofuran,acetonitrile and dichloromethane. In a further embodiment, thedeprotection is carried out a temperature from about 20° C. to about 50°C., e.g., about 37° C. to about 42° C. In certain embodiments, thereaction mixture is filtered through a pad of celite and the solvent(s)is removed by evaporation. In certain embodiments, amine f is used inthe next step without additional purification.

In reaction 6, the amine f is alkylated with RX to give a quaternaryammonium compound of Formula A. In certain embodiments, R is lower alkyland X is halo. In a further embodiment, the halo is iodide. In a furtherembodiment, RX is methyl iodide, i.e., the compound of Formula A is atrimethylammonium compound. In certain embodiments, the alkylation iscarried out in the presence of a base, such as potassium carbonate. Incertain embodiments, the alkylation is carried out in a solvent selectedfrom the group consisting of C₁-C₄ alcohol (e.g., methanol),tetrahydrofuran, acetonitrile and dichloromethane. In certainembodiments, the alkylation is carried out a temperature from about 20°C. to about 50° C., e.g., about 37° C. to about 42° C. In certainembodiments, the reaction mixture is filtered and the solvent is removedby evaporation to give a compound of Formula A. In certain embodiments,a compound of Formula A is purified by crystallization. In certainembodiments, a compound of Formula A is dissolved in a solvent selectedfrom the group consisting of C₁-C₄ alcohol (e.g., methanol),tetrahydrofuran, acetonitrile and dichloromethane to give a solution anda solvent, i.e., an anti-solvent, selected from the group consisting ofacetone, methyl ethyl ketone and ethyl acetate is added to inducecrystallization. In further embodiments, the anti-solvent is ethylacetate. The quaternary ammonium compound can form a pharmaceuticallyacceptable salt with any suitable pharmaceutically acceptable anion, X⁻,to provide a compound of Formula A.

Scheme 2 depicts the synthesis of compounds of Formula B wherein R isselected from the group consisting of hydrogen and lower alkyl, R¹ isselected from the group consisting of hydrogen, optionally substitutedalkyl, aralkyl, optionally substituted cycloalkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted aryl, optionally substituted heteroaryl and optionallysubstituted heterocyclo, R^(2b) and R^(3b) are independently selectedfrom the group consisting of hydrogen and monovalent pharmaceuticallyacceptable cation, or taken together R^(2b) and R^(3b) represent adivalent pharmaceutically acceptable cation, and X⁻ is apharmaceutically acceptable anion, or X⁻ and R^(2b) are absent and thecompound of Formula B is a zwitterion, and R⁴ is selected from the groupconsisting of hydrogen, optionally substituted alkyl and COR⁵.

In Scheme 2, R^(2a) and R^(3a) are removed from a compound of Formula Ato give a compound of Formula B. Compounds of Formula B wherein R^(2b)and R^(3b) are hydrogen may be prepared from phosphonate esters usingknown cleavage methods. For example, silyl halides are generally used tocleave various phosphonate esters and give the desired phosphonic acidupon mild hydrolysis of the resulting silyl phosphonate esters. Whenneeded, acid scavengers (for example, HMDS) can be used for the acidsensitive compounds. Such silyl halides include TMSCl (J. Org. Chem.28:2975 (1963)), TMSBr (Tetrahedron Lett. 155 (1977)) and TMSI (J. Chem.Soc., Chem. Commu. 870 (1978)). Alternatively, phosphonate esters can becleaved under strong acid conditions (Tetrahedron Lett. 33:4137 (1992);Synthesis-Stuttgart 10:955 (1993)). Those phosphonate esters can also becleaved via dichlorophosphonates prepared by treating the phosphonateesters with halogenating agents such as PCl₅, SOCl₂ and BF₃ (J. Chem.Soc. 238 (1961)) followed by aqueous hydrolysis to give the phosphonicacids. Aryl and benzyl phosphonate esters can be cleaved under hydrogeno lysis conditions (Synthesis 412 (1982); J. Med. Chem. 28:1208(1985)) or metal reduction conditions (J. Chem. Soc. 99:5118 (1977)).Electrochemical (J. Org. Chem. 44:4508 (1979)) and pyrolysis (Synth.Commu. 10:299 (1980)) conditions have also been used to cleave variousphosphonate esters.

Thus, in one embodiment, R^(2a) and R^(3a) are benzyl. In a furtherembodiment, the benzyl groups are removed under an atmosphere ofhydrogen gas and palladium on carbon is the catalyst. In certainembodiments the benzyl groups are removed in a solvent selected from thegroup consisting of C₁-C₄ alcohol (e.g., methanol), tetrahydrofuran,acetonitrile and dichloromethane. In certain embodiments, the benzylgroups are removed at temperature from about 20° C. to about 50° C.,e.g., about 37° C. to about 42° C. In certain embodiments, the reactionmixture is filtered through a pad of celite and the solvent(s) areremoved by evaporation to give a compound of Formula B wherein R^(2b)and R^(3b) are hydrogen.

In additional embodiments, R^(2a) and R^(3a) of a compound of FormulaIII, or a stereoisomer or mixture of stereoisomers thereof, are methyl.In certain embodiments, the methyl groups are removed using bromotrimethylsilane. In one embodiment the methyl groups are removed in asolvent selected from the group consisting of tetrahydrofuran anddichloromethane. In additional embodiments, the methyl groups areremoved at temperature from about 20° C. to about 50° C., e.g., fromabout 37° C. to about 42° C. In additional embodiments, the solvents areremoved by evaporation. In additional embodiments, the reaction mixtureis dissolved in water. In additional embodiments, the reaction mixtureis filtered and the solvent(s) are removed by evaporation to give acompound of Formula B, or a stereoisomer or mixture of stereoisomersthereof, wherein R^(2b) and R^(3b) are hydrogen.

In certain embodiments, a compound of Formula B is purified bycrystallization. In certain embodiments, a compound of Formula A isdissolved in a solvent selected from the group consisting of C₁-C₄alcohol (e.g., methanol), tetrahydrofuran, and acetonitrile to give asolution and a solvent, i.e., an anti-solvent, selected from the groupconsisting of acetone, methyl ethyl ketone and ethyl acetate is added toinduce crystallization. In further embodiments, the anti-solvent isethyl acetate. The quaternary ammonium compound can form apharmaceutically acceptable salt with any suitable pharmaceuticallyacceptable anion, X⁻, to provide a compound of Formula B.

In Scheme 3, a compound of Formula C is prepared as described above inScheme 1, starting from an amino acid g, i.e., a L-amino acid, e.g.,L-leucine.

In Scheme 4, the diastereomers of a compound of Formula C are separatedinto compounds of Formula D, i.e., the 2R, 3S-isomer, and Formula E,i.e., the 2S, 3S-isomer. In certain embodiments, the diastereomers of acompound of Formula C are separated by crystallization. In certainembodiments, the crystallization comprises: (a) dissolving a compound ofFormula C in a solvent or solvent system, i.e., a mixture of solvents,e.g., methanol/water, ethanol/water, tetrahydrofuran/water,acetonitrile/water, etc., to give a solution; (b) allowing precipitationto occur; and (c) separating crystalline product from said solution. Incertain embodiments, the solution is a homogeneous solution. In certainembodiments, the solvent or solvent system is selected from the groupconsisting of dichloromethane, methanol, methanol/water, ethanol,ethanol/water, isopropanol, isopropanol/water, tetrahydrofuran,tetrahydrofuran/water, acetonitrile and acetonitrile/water. In certainembodiments, precipitation of the desired product, i.e., a compound ofFormulae D or E, is induced by adding an anti-solvent. In furtherembodiments, the anti solvent is selected from the group consisting ofhexane, ethyl acetate, acetone, methyl ethyl ketone and methyl t-butylether, particularly ethyl acetate.

In additional embodiments, precipitation of the product duringcrystallization is induced by cooling the solution. In a furtherembodiment, the solution is cooled to about 10° C., to about 5° C. or toabout 0° C.

In additional embodiments, the solution is heated before the addition ofthe anti-solvent, during the addition of the anti-solvent or after theaddition of the anti-solvent.

In certain embodiments, the crystalline product is isolated byfiltration.

In Scheme 5, a compound of Formula F is prepared as described above inScheme 2, starting from a compound of Formula D. In a similar fashion, acompound of Formula G is prepared from a compound of Formula E.

In Scheme 6, a compound of Formula H is prepared as described above inScheme 1, starting from an amino acid h, i.e., a D-amino acid, e.g.,D-leucine.

In Scheme 7, a compound of Formula K, i.e., the 2R, 3R-isomer, andFormula L, i.e., the 2S, 3R-isomer, are prepared as described above inScheme 4, starting from a compound of Formula J.

In Scheme 8, a compound of Formula M is prepared as described above inScheme 2, starting from a compound of Formula K. In a similar fashion, acompound of Formula N is prepared from a compound of Formula L.

Scheme 9 depicts the synthesis of β-amino-γ-aminophosphonate andβ-amino-γ-aminophosphonate analogs of Formula O. In reaction 1, the oxogroup, i.e., C═O, of a β-keto-γ-aminophosphonate a is converted toβ-amino-γ-aminophosphonate b via reductive amination. In one embodiment,a is reacted with H₂NR¹⁰ in the presence of a reducing agent, such as,but not limited to, NaCNBH₃ or NaBH(OAc)₃, in an organic solvent suchas, but not limited to, tetrahydrofuran. A compound of Formula O can beprepared from b using the methods described in Scheme 1 and routineamine protection/deprotection strategies well-known to one of ordinaryskill in the art.

It will be readily apparent to one of ordinary skill in the relevantarts that other suitable modifications and adaptations to the methodsand applications described herein may be made without departing from thescope of the invention or any embodiment thereof. It is to be understoodthat while the invention has been described in conjunction with thedetailed description thereof, the foregoing description is intended toillustrate and not limit the scope of the invention, which is defined bythe scope of the appended claims. Other aspects, advantages, andmodifications are within the scope of the claims. Having now describedthe present invention in detail, the same will be more clearlyunderstood by reference to the following examples, which are includedherewith for purposes of illustration only and are not intended to belimiting of the invention.

Example 1 Synthesis of(2R,3S)-(2-hydroxy-5-methyl-3-trimethylamonium-hexyl)-phosphonic aciddimethyl ester; iodide

Scheme 10 depicts the synthesis of(2R,3S)-(2-hydroxy-5-methyl-3-trimethylamonium-hexyl)-phosphonic aciddimethyl ester; iodide.

Reaction 1: 2.5 g of L-leucine (19.05 mmol) was taken up in methanol (10ml) and K₂CO₃ (32.40 mmol) was added. The mixture was stirred and benzylbromide (62.89 mmol) was slowly added. The reaction mixture was heatedunder reflux for 24 h. The solvent was removed under reduced pressure.Ethyl acetate was added, the salts were removed by filtration and thefiltrate was concentrated under reduced pressure to give compound 2. Theresidual oil was dissolved in anhydrous THF (12 ml) (solution A).

Reaction 2: A solution of dimethylmethylphosphonate (60.98 mmol) inanhydrous THF (12 ml) was cooled at −50° C. and a solution of n-BuLi inhexane (2.4 M, 67.08 mmol) was slowly added. The resulting solution wasstirred at −50° C. for 1 h and added to solution A from reaction 1above. The reaction mixture was stirred at −50° C. for 3 h before theaddition of solution of HCl 5% (v/v). The reaction mixture wasconcentrated under reduced pressure. The residual oil was dissolved inethyl acetate, and washed with 10 ml portions of water. The organicphase was dried over anhydrous Na₂SO₄ and filtered. The solvent wasevaporated to give compound 3 as a crude oil.

Reaction 3: A mixture of the crude oil(3-dibenzylamino-5-methyl-2-oxohexyl)-phosphonic acid dimethyl ester,i.e., compound 3, obtained in reaction 2, was dissolved in about 10%methanol and about 90% THF (v/v) and cooled at −10° C. NaBH₄ (19.05mmol) was added to the mixture in small portions with vigorous stirring.The mixture was stirred at temperature of −10 to 5° C. for 4 h and HCl5% (v/v) was added. The reaction mixture was concentrated under reducedpressure. The residual oil was dissolved in ethyl acetate, and washedwith 10 ml portions of water. The organic phase was dried over anhydrousNa₂SO₄ and filtered. The solvent was evaporated off to leave(3-dibenzylamino-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethylester, compound 4, as a crude oil. ³¹P NMR (at 200 MHz, CDCl₃) indicatedan 80:20 ratio of 2R,3S (δ 34.53) to 2S,3S (δ 35.15) isomers.(3-Dibenzylamino-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethylester can be isolated by chromatography or used in the nexthydrogenolysis step directly without isolation.

Reaction 4: A diastereomeric mixture of the crude(3-dibenzylamino-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethylester, compound 4, obtained in reaction 3, was treated withpalladium-carbon (2.0% wt) as the catalyst in methanol (40 mL). Themixture reaction was stirred for 12 h under a hydrogen gas atmosphere at37-42° C., and after this period of time, the mixture was filteredthrough a pad of Celite, and the solvents were removed under reducedpressure to leave (3-amino-2-hydroxy-5-methyl-hexyl)-phosphonic aciddimethyl ester, compound 5, as a crude oil.

Reaction 5: A mixture of the crude(3-amino-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethyl ester, i.e.,compound 5, obtained in reaction 4 was dissolved in methanol (30 mL) andK₂CO₃ (33.38 mmol) and CH₃I (66.67 mmol) were added. The reactionmixture was stirred for 12 h at 34-37° C., and after this period of timewas filtered and the solvent was removed under reduced pressure off toleave (3-trimethylamonium-2-hydroxy-5-methyl-hexyl)-phosphonic aciddimethyl ester; iodide, compound 6, as a crude oil. The diasteromericmixture was analyzed by ³¹P NMR (at 200 MHz, CDCl₃), to give compound ofFormula 2R,3S (δ 31.35) and 2S,3S (δ 30.91) in the ratio 76:24,respectively.

Reaction 6: The (3-trimethylamonium-2-hydroxy-5-methyl-hexyl)-phosphonicacid dimethyl ester; iodide, i.e., compound 6, from reaction 5 wasdissolved in methanol. Crystallization was induced by the addition ofethyl acetate. The product that crystallized was filtered and driedunder reduced pressure to give 2.1 g (26.93%) of the title compound.

¹H NMR (400 MHz, CDCl₃) δ 1.06 (d, J=6.8 Hz, 3H, (CH₃)₂CH), 1.08 (d,J=6.4 Hz, 3H, (CH₃)₂CH), 1.50 (dd, J=13.6, 9.2 Hz, 1H, CH₂CH), 1.73 (m,1H, CH(CH₃)₂) 1.89 (ddd, J=13.6, 8.8, 4.2 Hz, 1H, CH₂CH), 2.49 (ddd,J=18.6, 15.6, 8.6 Hz, 1H, CH₂P), 2.63 (ddd, J=19.2, 15.6, 4.8 Hz, 1H,CH₂P), 3.45 (s, 9H, (CH₃)₃N), 3.78 (d, J=10.8 Hz, 3H, (CH₃))₂P), 3.80(d, J=10.8 Hz, 3H, (CH₃O)₂P), 3.99 (d, J=8.8 Hz, 1H, CHN), 4.43 (m, 1H,CHOH).

¹³C NMR (100 MHz, CDCl₃) δ 21.95 ((CH₃)₂CH), 23.76 ((CH₃)₂CH), 26.78(CH(CH₃)₂), 32.47 (d, J=135.1 Hz, CH₂P), 38.03 (CH₂CH), 53.35((CH₃O)₂P), 54.12 ((CH₃)₃N), 67.30 (CHN), 73.63 (CHOH). ³¹P NMR (200MHz, CDCl₃) δ 31.31.

Example 2 Synthesis of(2R,3S)-(2-hydroxy-5-methyl-3-trimethylamonium-hexyl)-phosphonic acid;iodide

Scheme 11 depicts the synthesis of(2R,3S)-(2-hydroxy-5-methyl-3-trimethylamonium-hexyl)-phosphonic acid;iodide.

Reaction 1: 10 g of L-leucine (76.23 mmol) was taken up in methanol (40ml) and K₂CO₃ (129.60 mmol) was added. The mixture was stirred andbenzyl bromide (251.56 mmol) was slowly added. The reaction mixture washeated under reflux for 24 h. The solvent was removed under reducedpressure. Ethyl acetate was added, the salts were removed by filtrationand the filtrate was concentrated under reduced pressure to givecompound 2. The residual oil was dissolved in anhydrous THF (45 ml)(solution A).

Reaction 2: A solution of dimethylmethylphosphonate (243.92 mmol) inanhydrous THF (45 ml) was cooled at −50° C. and a solution of n-BuLi inhexane (2.4 M, 268.32 mmol) was slowly added. The resulting solution wasstirred at −50° C. for 1.5 h and added to solution A from reaction 1above. The reaction mixture was stirred at −50° C. for 3 h before theaddition of solution of HCl 5% (v/v). The reaction mixture wasconcentrated under reduced pressure. The residual oil was dissolved inethyl acetate, and washed with 30 ml portions of water. The organicphase was dried over anhydrous Na₂SO₄ and filtered. The solvent wasevaporated to give compound 3 as a crude oil.

Reaction 3: A mixture of the crude oil(3-dibenzylamino-5-methyl-2-oxohexyl)-phosphonic acid dimethyl ester,i.e., compound 3, obtained in reaction 2, was dissolved in about 10%methanol and about 90% THF (v/v) and cooled at −10° C. NaBH₄ (68.58mmol) was added to the mixture in small portions with vigorous stirring.The mixture was stirred at temperature of −10 to 5° C. for 4 h and HCl5% (v/v) was added. The reaction mixture was concentrated under reducedpressure. The residual oil was dissolved in ethyl acetate, and washedwith 30 ml portions of water. The organic phase was dried over anhydrousNa₂SO₄ and filtered. The solvent was evaporated off to leave(3-dibenzylamino-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethylester as a crude oil. ³¹P NMR (at 200 MHz, CDCl₃) indicated an 80:20ratio of 2R,3S (δ 34.53) to 2S,3S (δ 35.15) isomers. The diastereomericmixture was purified by flash chromatography on silica gel usinghexane/ethyl acetate as the eluent to afford compound 2R,3S-4 (20.753 g;64.9% yield; ³¹P NMR at 200 MHz, CDCl₃; δ 34.61).

Reaction 4: 10.365 g of(2R,3S)-3-dibenzylamino-2-hydroxy-5-methyl-hexyl)-phosphonic aciddimethyl ester, compound 2R,3S-4, obtained in reaction 3, was treatedwith palladium-carbon (2.0% wt) as the catalyst in methanol (40 mL). Themixture reaction was stirred for 12 h under a hydrogen gas atmosphere at37-42° C., and after this period of time, the mixture was filteredthrough a pad of Celite, and the solvents were removed under reducedpressure to leave (2R,3S)-3-amino-2-hydroxy-5-methyl-hexyl)-phosphonicacid dimethyl ester, compound 2R,3S-5, as a crude oil.

Reaction 5: The crude(2R,3S)-3-amino-2-hydroxy-5-methyl-hexyl)-phosphonic acid dimethylester, i.e., compound 2R,3S-5, obtained in reaction 4 was dissolved inmethanol (30 mL) and K₂CO₃ (43.26 mmol) and CH₃I (86.52mmol) were added.The reaction mixture was stirred for 12 h at 32-35° C., and after thisperiod of time was filtered and the solvent was removed under reducedpressure off to leave(2R,3S)-(3-trimethylamonium-2-hydroxy-5-methyl-hexyl)-phosphonic aciddimethyl ester; iodide, compound 2R,3S-6, as a crude oil. ³¹P NMR (at200 MHz, CDCl₃) indicated that the product had 2R,3S stereochemistry (δ31.23).

Reaction 6: The crude(2R,3S)-3-trimethylamonium-2-hydroxy-5-methyl-hexyl)-phosphonic aciddimethyl ester; iodide, i.e., compound 2R,3S-6, obtained in reaction 5was treated with bromotrimethylsilane (54.39 mmol) in dichloromethaneunder nitrogen atmosphere. The mixture reaction was stirred for 4 h at37-42° C., and after this period of time the volatile materials wereevaporated under reduced pressure, water was then added. After 2 h thesolvent were remover under reduced pressure off to leave(2R,3S)-3-trimethylamonium-2-hydroxy-5-methyl-hexyl)-phosphonic acid;iodide, compound 7. Compound 7 was dissolved in methanol andcrystallization was induced by the addition of ethyl acetate. Theproduct that crystallized was filtered and dried under reduced pressureto give 5.8 g (61.63%) of the title compound. ¹H NMR (200 MHz, CD₃OD) δ1.04 (d, J=6.6 Hz, 6H, (CH₃)₂CH), 1.73 (m, 3H, CH₂CH, CH(CH₃)₂), 2.29(m, 2H, CH₂P), 3.28 (s, 9H, (CH₃)₃N), 3.57 (d, J=8.2 Hz, 1H, CHN), 4.33(m, 1H, CHOH). ¹³C NMR (50 MHz, CD₃OD) δ 21.64 ((CH₃)₂CH), 23.92((CH₃)₂CH), 27.43 (CH(CH₃)₂), 36.48 (d, J=134.1 Hz, CH₂P), 38.52(CH₂CH), 53.98 ((CH₃)₃N), 68.89 (CHN), 73.48 (d, J=11.0 Hz, CHOH).

³¹P NMR (81 MHz, CD₃OD₃) δ 24.898.

Example 3 Induction of Changes of Utilization of Glucose in HepaticCells

WRL-68 cells were thawed and maintained in culture with MinimalEssential Medium (Gibco BRL) containing 4 mM Glutamine, 1% non essentialaminoacids (Gibco BRL), 10% Fetal Calf Serum (FCS), and 100 μg/ml ofampicillin. WRL-68 cells were seeded at 2×10⁶ cells/cm² in completemedia in 75 cm² cell culture flasks, cells were maintained for 24 hoursat 37° C. under an atmosphere of 5% CO2. The medium was changed after 24hr. Cells were passaged by trypsinization using a 0.025% trypsinsolution containing 0.01% N,N,-di-ethyldithiocarbamic acid sodium salt(EDTA). In order to evaluate the effect of analog of carnitine onutilization of glucose levels a model of hyperglycemia in vitro(Nakajina et al., J. Biol. Chem. 275:20880-20886 (2000)) was developed.Two days after plating, the medium was changed to F-12K containing 7 mMD-glucose and 10% fetal bovine serum, and the culture was continued for2 more days. The cells were then cultured in serum-free F-12K mediumcontaining 30 mM D-glucose for 24 h. Cells were treated with thediastereomeric mixture of compound 7 at concentrations of 0.01, 0.1,1,10, 100 μM, and 1 mM and 2 mM during 24 hours. Samples of conditionedmedia and cell lysates were kept to minus 8° C. Glucose and glycogenlevels were measured by using a Glucose PAP-SL Kit (Tech, USA) andglycogen assay kit (Biovision Research Products, California, USA).

The diastereomeric mixture of compound 7 in concentrations of 1, 10 and100 μM reduced the extracellular glucose levels in 16%, 18% and 23%,respectively; while a reduction of 30 and 32% was observed withconcentrations of 1 and 2 mM (FIG. 1). No changes in cellular glucosewere observed with any concentration (FIG. 2). An increase in cellularglycogen levels were observed with diastereomeric mixture of compound 7at a concentration of 2 mM (FIG. 3).

Example 4 Safety of Analog of Carnitine in Human Cells and Evaluation ofTeratogenicity

Two cell lines were used for this study: 293Q cells derived from normalepithelial cells of human fetal kidney (CRL-1573 ATCC) and WRL-68 cellsderived from epithelial cells of human liver (CRL-48 ATCC). Cell lineswere cultured in minimal essential medium (MEM, GIBCO BRL Inc., GrandIsland, N.Y.), supplemented with nonessential amino acids (GIBCO BRLInc., Grand Island, N.Y.), 10% fetal calf serum (GIBCO BRL Inc., GrandIsland, N.Y.), 1-glutamine (2 mol/L), and antibiotics. Cells were platedin 100-mm culture dishes (10⁶ cells/dish), and maintained at 37° C.under an atmosphere of 5% CO₂ in humidified air. Subcultures wereobtained by trypsinization (0.025% trypsin solution containing 0.01%N,N-diethyldithiocarbamic acid sodium salt, EDTA). For cytologicalinvestigation, 10⁵ cells per mL of MEM medium were used. Cells weretreated with the following concentrations of the diastereomeric mixtureof compound 7: 0.01, 0.1, 1, 10, and 100 μM, and 1 and 2 mM. Afterincubation of the cells with the extract, they were collected forfurther cytological investigations.

Cells were incubated in 96-well plates. After 24 h, the medium wasremoved; the cells were washed twice with PBS and then incubated withanalog. After 24 h, both cells and conditioned media were collected andprocessed. Cell viability was measured by a3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay(Wang et al., Journal Ocular. Pharm. Ther. 12: 35-43 (1996)). Briefly,20 μL MTT (5 g/L) was added to each well and incubated with the culturefor an additional 4 h at 37° C., 5% CO₂ and then culture media wasdiscarded followed by addition of 200 μL DMSO with 25 μL Sørensen'sglycine buffer (glycine 0.1 M, NaCl 0.1 M, pH 10.5) to each well. Whenthe blue crystals were dissolved, the optical density was determined ona microplate reader at 450 nm. In order to evaluate the effect of thediastereomeric mixture of compound 7 on cell proliferation the MTT wasalso used. To this end cells were collected everyday during four daysfor hepatic cells and seven days for renal cells. Cells exposed to thediastereomeric mixture of compound 7 did not show citotoxicity oreffects on cell proliferation (FIGS. 4, 5 and 6).

Example 5 Teratogenicity Assay

A teratogenicity assay was carried out as described by Jelinek et al.,Func. Devel. Morph. 4:317-23 (1994). Fertile White Leghorn chicken eggswere obtained from A.L.P.E. S.A. (Puebla, Mexico) and were stored at 6°C. Sixty fertilized eggs were weighed, sterilized, and divided into sixgroups. First group served as a non-treated control. The next three wereexposed to the diastereomeric mixture of compound 7 (1, 10, and 100 μM).The last group received caffeine (10 mg/mL) and as positive control.Test solutions (1 mL) were added to the air sac under sterileconditions. Each solution was injected after drilling into the shell atthe blunt end of the egg; after injection, the holes were immediatelysealed with melted paraffin wax. The eggs were then transferred andmaintained in a forced draft incubator at 37.5° C. with a relativehumidity of 55% until the desired stage of development was reached.

To determine the concentration dependency of the diastereomeric mixtureof compound 7, a histological analysis was carried out. Embryos in eachgroup were fixed in buffered formal saline (pH 7.4), dehydrated, andembedded in paraffin blocks. Paraffin tissue sections of 6 μm werestained with acetocarmine for routine histological examination. Theembryo was examined and staged according to morphological criteriapreviously outlined by Hamburger et al. (Hamburger et al., J. Morph.88:49-921951 (1951). Embryonic stages at the time of the diastereomericmixture of compound 7 application varied from 14-16, which correspondapproximately to developed somites numbered 22-28. No alterations inembryos treated with the diastereomeric mixture of compound 7 were found(FIG. 7 and Table 1).

TABLE 1 Teratogenic evaluation of diastereometic mixturecompound 7Embryonic region affected 48 h of Axial exposition skeleton CNSVasculature Heart Somites Deaths Ringer¹ 0/10 0/10 0/10 0/10 0/10 0/10Cpd 7 1 μM 0/10 0/10 0/10 0/10 0/10 0/10 Cpd 7 100 μM 0/10 0/10 0/100/10 0/10 0/10 Cpd 7 1 mM 0/10 0/10 0/10 0/10 0/10 0/10 Caffeine² 10/10 8/10 6/10 8/10 8/10 2/10 ¹Negative control; ²Positive control; Thefractions represent the number of abnormal embryos and the totalexamined for each developmental region of the embro.

The present invention has been described with reference to certainembodiments thereof. However, the scope of the invention is not limitedto the embodiments described or exemplified. Workers of ordinary skillin the relevant arts will readily appreciate that other embodiments andexamples can be practiced without departing from the scope of thepresent invention. All such variations are considered to be part of, andtherefore encompassed by, the present invention.

All publications, patents and patent applications mentioned orreferenced in this specification are herein incorporated by reference tothe same extent as if each individual publication, patent or patentapplication was specifically and individually indicated to beincorporated by reference.

1. A compound of Formula I

wherein: R is selected from the group consisting of hydrogen and loweralkyl; R¹ is selected from the group consisting of hydrogen, optionallysubstituted alkyl, aralkyl, optionally substituted cycloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, optionally substituted heteroaryl, andoptionally substituted heterocyclo; R² and R³ are independently selectedfrom the group consisting of hydrogen, optionally substituted alkyl,aralkyl, optionally substituted aryl, and monovalent pharmaceuticallyacceptable cation, or taken together R² and R³ represent a divalentpharmaceutically acceptable cation; R⁴ is selected from the groupconsisting of hydrogen, optionally substituted alkyl, optionallysubstituted cycloalkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, and COR⁵; R⁵ is selected from the group consistingof optionally substituted alkyl, aralkyl, optionally substitutedcycloalkyl, optionally substituted alkenyl, optionally substitutedalkynyl, optionally substituted aryl, optionally substituted heteroaryl,and optionally substituted heterocyclo; X⁻ is a pharmaceuticallyacceptable anion, or X⁻ and R² are absent and the compound of Formula Iis a zwitterion; Z is selected from the group consisting of O and NR¹⁰;and R¹⁰ is selected from the group consisting of hydrogen, optionallysubstituted alkyl, aralkyl, optionally substituted cycloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, optionally substituted heteroaryl, andoptionally substituted heterocyclo; or a pharmaceutically acceptablehydrate, crystalline form or amorphous form thereof, or a stereoisomeror mixture of stereoisomers thereof.
 2. (canceled)
 3. The compound ofclaim 1, wherein R is methyl. 4.-5. (canceled)
 6. The compound of claim1, wherein R¹ is isobutyl. 7.-11. (canceled)
 12. The compound of claim1, wherein Z is O.
 13. (canceled)
 14. The compound of claim 12, whereinR⁴ is hydrogen. 15.-16. (canceled)
 17. The compound of claim 1, whereinR² and R³ are hydrogen.
 18. The compound of claim 1, wherein X⁻ isselected from the group consisting of hydroxide, chloride, bromide,iodide, sulphate, nitrate, phosphate, formate, acetate, maleate,fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate,lactate, gluconate, trifluoroacetate, methanesulphonate, besylate andp-toluenesulphonate.
 19. (canceled)
 20. The compound of claim 3, whereinR¹ is selected from the group consisting of methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl, and phenyl,R² and R³ are independently selected from the group consisting hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, isopenyl, benzyl, and phenyl, and R⁴ and R¹⁰ are hydrogen.21.-23. (canceled)
 24. The compound of claim 1, wherein said compound ofFormula I is in the 2R, 3S-isomeric form. 25.-38. (canceled)
 39. Thecompound of claim 1, wherein said compound is:


40. A process for preparing a compound of Formula III:

wherein: R is selected from the group consisting of hydrogen and loweralkyl; R¹ is selected from the group consisting of hydrogen, optionallysubstituted alkyl, aralkyl, optionally substituted cycloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, optionally substituted heteroaryl andoptionally substituted heterocyclo; R^(2a) and R^(3a) are independentlyselected from the group consisting of optionally substituted alkyl,aralkyl and optionally substituted aryl; and X⁻ is a pharmaceuticallyacceptable anion; said method comprising: (a) reacting a compound ofFormula IV:

with RX, to give said compound of Formula III, and (b) isolating saidcompound of Formula III.
 41. The process of claim 40, wherein saidcompound of Formula III is the 3S-isomer and said compound of Formula IVis the 3S-isomer. 42.-54. (canceled)
 55. The process of claim 41 furthercomprising: (c) isolating the 2S,3S-isomer substantially free from the2R,3S-isomer, or (d) isolating the 2R,3S-isomer substantially free fromthe 2S,3S-isomer. 56.-63. (canceled)
 64. The process of claim 55 furthercomprising: (e) removing R^(2a) and R^(3a) from said 2S,3S-isomer, togive a compound of Formula 2S,3S-X:

or, (f) removing R^(2a) and R^(3a) from said 2R,3S-isomer, to give acompound of Formula 2R,3S-X:

wherein R^(2b) and R^(3b) are selected from the group consisting ofhydrogen and monovalent pharmaceutically acceptable cation, or takentogether R^(2b) and R^(3b) represent a divalent pharmaceuticallyacceptable cation, or X⁻ and R^(2b) are absent.
 65. The process of claim40, wherein said process comprises removing R⁶, or R⁶ and R⁷, from acompound of Formula V:

wherein: R⁶ is an amine protecting group; and R⁷ is selected from thegroup consisting of hydrogen and amine protecting group, or R⁶ and R⁷taken together represent an amine protecting group, to give saidcompound of Formula IV. 66.-67. (canceled)
 68. The process of claim 65,wherein said process comprises reduction of a compound of Formula VI:

to give said compound of Formula V. 69.-77. (canceled)
 78. The processof claim 68, wherein said process comprises condensing a compound ofFormula VII:

with a compound of Formula VIII:

to give said compound of Formula VI, wherein: R⁸ is selected from thegroup consisting of optionally substituted alkyl, aralkyl, andoptionally substituted aryl. 79.-84. (canceled)
 85. The process of claim78, wherein said process comprises condensing a compound of Formula IX:

with LiR⁹, to give said compound of Formula VIII, wherein R⁹ is selectedfrom the group consisting of lower alkyl and aryl. 86.-89. (canceled)90. The process of claim 78, wherein said process comprises: (a)protecting the amine; and (b) esterifying the carboxylic acid of anamino acid of Formula XI:

to give said compound of Formula VII. 91.-99. (canceled)
 100. Theprocess of claim 40, wherein said compound of Formula III is obtainedvia a one pot process from a compound of Formula XI:

101.-109. (canceled)
 110. A pharmaceutical composition comprising thecompound of claim 1 and a pharmaceutically acceptable carrier. 111.-114.(canceled)
 115. A method for inhibiting carnitine acyltransferase in acell comprising contacting said cell with the compound of claim 1.116.-117. (canceled)
 118. A method of treating, ameliorating, orpreventing a disorder or condition responsive to the inhibition ofcarnitine acyltransferase in a patient in need thereof, comprisingadministering to said patient a therapeutically effective amount of thecompound of claim
 1. 119.-123. (canceled)
 124. The method of claim 118,wherein said disorder or condition is associated with disorders in fattyacid metabolism.
 125. The method of claim 124, wherein said disorder orcondition is selected from the group consisting of non-insulin dependentdiabetes mellitus, obesity, hyperlipoproteinemia, hyperlipidemia,myocardial dysfunction, renal anemia and Alzheimer's disease.
 126. Themethod of claim 125, wherein said disorder or condition is non-insulindependent diabetes.
 127. The method of claim 125, wherein said disorderor condition is obesity.